Transglutaminase gene products

ABSTRACT

The invention provides a nucleotide sequence comprising at least a portion of the nucleotide sequence of FIG.  10 A, FIG.  6 B or FIG.  10 A or FIG.  10 B; nucleotides which hybridise to the nucleotide sequences of FIG.  6 A, FIG.  6 B or FIG.  10 A or FIG.  10 B; nucleotides which are degenerate to the nucleotide sequences of FIG.  6 A, FIG.  6 B or FIG.  10 A or FIG.  10 B; all of which nucleotides encode a polypeptide having transglutaminase activity.

[0001] The present invention relates to the identification of noveltransglutaminase enzymes TG_(Z) and TG_(Y).

[0002] Transglutaminases are a family of structurally and functionallyrelated enzymes that catalyze the post-translational modification ofproteins via a Ca²⁺ dependant transferase reaction between theγ-carboxamide group of a peptide-bound glutamine residue and variousprimary amines. Most commonly, γ-glutamyl-ε-lysine cross links areformed in or between proteins by reaction with the ε-amino group oflysine residues. Analysis of the three-dimensional structure of thea-subunit of factor XIII showed that transglutaminases contain a centralcore domain containing enzymatic activity, and a N-terminal β-sandwichdomain and two C-terminal β-barrel domains, which are thought to beinvolved in the regulation of enzyme activity and specificity.

[0003] Seven different transglutaminase genes have been characterised inhigher vertebrates on the basis of their primary structure (Aeschlimann,D, and Paulsson, M (1994) Thromb. Haemostasis 71: 402-415 Aeschlimann etal: (1998) J. Biol Chem 273, 3542). Transglutaminases can be foundthroughout the body, but each transglutaminase is characterised by itsown typical tissue distribution, although each may be present in anumber of different tissue types often in combination with othertransglutaminases. Transglutaminase gene products have specificfunctions in the cross linking of particular proteins or tissuestructures. For review see Aeschlimann and Paulsson (1994) (supra) andAeschlimann and Tholmazy (2000) Connective Tissue Res. 41, 1-27. Forexample, factor XIIIa stabilises the fibrin clot in haemostasis, whereasprostate transglutaminase (TG_(P))¹ is involved in semen coagulation.Other transglutaminases have adopted additional functions such as thetissue transglutaminase (TG_(C)), which is involved in GTP-binding inreceptor signalling, and band 4.2 protein which functions as astructural component of the cytoskeleton. Four transglutaminases havebeen shown to be expressed during the different stages of epidermalgrowth and differentiation. Three of these, keratinocytetransglutaminase (TG_(K)), epidermal transglutaminase (TG_(E)) andTG_(X), are associated with keratinocyte terminal differentiation andthe cross-linking of structural proteins to form the cornified envelope.The fourth enzyme TG_(C), is expressed in skin primarily in the basalcell layer, and plays a role in the stabilisation of the dermo-epidermaljunction. The importance of proper cross-linking of the cornifiedenvelope is exemplified by the pathology seen in patients suffering froma severe form of the skin disease referred to as congenital ichthyosis,which has been linked to mutations in the gene encoding TG_(K).

[0004] All transglutaminase enzymes appear to be encoded by a family ofclosely related genes. Alignment of these genes demonstrates that allmembers of the transglutaminase family exhibit a similar geneorganisation, with remarkable conservation of intron distribution.Furthermore, phylogenetic analysis indicates that an early geneduplication event subsequently gave rise to two differenttransglutaminase lineages; one comprising TG_(C), TG_(E), and band 4.2protein; the other, factor XIIIa, TG_(K) and possibly also TG_(P)(Aeschlimann and Paulsson (1994) (supra)). The genes encoding TG_(K) andfactor XIIIa have been mapped to human chromosome 14q11.2 and chromosome6p24-25 respectively, whereas TG_(C) and TG_(E) have been mapped tochromosome 20q11, and TG_(P) has been mapped to chromosome 3p21-22.

[0005] Comparison of the structure of the individual transglutaminasegenes shows that they may be divided into two subclasses, wherein thegenes encoding TG_(C), TG_(E), TG_(P) and band 4.2 protein comprise 13exons and 12 introns, and the genes encoding factor XIIIa and TG_(K)contain two extra exons. Exon IX of the former group is separated intotwo exons (X and XI) in TG_(K) and factor XIIIa, and the amino-terminalextensions of TG_(K) and factor XIIIa comprise an additional exon.However, except for the acquisition of an additional intron and therecruitment of an exon by the genes encoding factor XIIIa and TG_(K),the gene structure is remarkably conserved among all members of thetransglutaminase gene family. Not only is the position of intron splicepoints highly conserved, but also the intron splice types. Thissimilarity in gene structure and homology of the primary structure ofthe transglutaminases provides further support for the proposition thatthe different transglutaminase genes are derived from a common ancestralgene.

[0006] The inventors have previously isolated a cDNA encoding a novelmember of the transglutaminase gene family TG_(X), from human foreskinkeratinocytes (Aeschilmann et al (1998) J. Biol. Chem., 273, 3452-3460).Two related transcripts with an apparent size of 2.2 and 2.8 kb wereobtained. The deduced amino acid sequence for the full-length geneproduct encodes a protein with 720 amino acids and a molecular mass of81 kDa. A sequence comparison of TG_(X) to the other members of thetransglutaminase gene family revealed that the domain structure and theresidues required for enzymatic activity and Ca²⁺ binding are conservedand show an overall sequence identity of about 35%, with the highestsimilarity being found within the enzyme's catalytic domain.

[0007] The inventors subsequently determined that TG_(X) is the productof a ˜35 kb gene located on chromosome 15, comprising 13 exons and 12introns. The intron splice sites were found to conform to the consensusfor splice junctions in eukaryotes. The transcription initiation site islocalised to a point 159 nucleotides upstream of the initiatormethionine and the likely polyadenylation site is localised ˜600nucleotides downstream of the stop codon. The two mRNA isoforms are theresult of alternative splicing of exon III and give rise to 2 proteinvariants of TG_(X) which comprise catalytic activity. TG_(X) isexpressed predominately in epithelial cells, and most prominently duringfoetal development, in epidermis and in the female reproductive system.

[0008] The inventors have now localised the TGM5 gene to chromosome15q15 by fluorescent in situ hybridisation. Band 4.2 protein haspreviously been mapped to this chromosomal region (Sung L. A. et al(1992) Blood 79: 2763-2770; Najfeld V. et al (1992) Am. J. Hum. Genet50: 71-75) and has subsequently been assigned to position 15q15.2 byexpression mapping of the LGMD2A locus on chromosome 15 (ChiannikulchaiN. et al (1995) Hum. Mol. Genet 4: 717-725). A short sequenceencompassing the left arm of one of the YAC clones (926G10²) used forexpression mapping matched with the sequence of intron 12 of the TGM5gene placing the genes encoding TG_(X) and band 4.2 protein in closeproximity on chromosome 15 (FIG. 5C). PCR with specific primers for 5′(exon I) or 3′ (exon XIII) sequences of band 4.2 protein as well assouthern blot analysis revealed that the BAC clones containing the TGM5gene also contained the EPB42 gene and that the 2 genes are arranged intandem.

[0009] Further analysis by the inventors has recently led to theidentification of two novel transglutaminase genes TGM7 and TGM6 whichencode the proteins TG_(Z) and TG_(Y) respectively. Alternative mRNAsequences of the TGM7 gene are given in FIG. 6A and FIG. 6B. The TGM7derived mRNA (FIG. 6A and FIG. 6B) comprises an open reading frame of2130 nucleotides and a polyadenylation signal (AATAAA) 158 nucleotidesdownstream of the termination codon (TGA). The deduced protein forTG_(Z) consists of 710 amino acids. The deduced protein for TG_(Z) fromFIG. 6A has a molecular mass of 79, 908 Da and an isoelectric point of6.7. The deduced protein for TG_(Z) from FIG. 6B has a molecular weightof 80,065 and an isoelectric point of 6.6.

[0010] The TGM6 full length transcript (FIG. 10A) comprises an openreading frame of 2109 nucleotides. The deduced protein for the long formof TG_(Y) consists of 708 amino acids and has a calculated molecularmass of 79, 466 Da and an isoelectric point of 6.9. The transcript forthe short form of TG_(Y) (FIG. 10B) comprises an open reading frame of1878 nucleotides and the deduced protein consists of 626 amino acidswith a molecular mass of 70, 617 Da and an isoelectric point of 7.6.

[0011] To analyse the relationship between the differenttransglutaminase genes, the inventors calculated their amino acidsimilarity based upon sequence alignments, and calculated theirevolutionary distances using different algorithms. All the algorithmsused predicted a close relationship between TG_(X), TG_(Z), TG_(Y),TG_(E), band 4.2 protein and TG_(C), and factor XIIIa and TG_(K),respectively. The grouping of TG_(X), TG_(Z), TG_(Y), TG_(E), TG_(C),and band 4.2 protein in one subclass and factor XIIIa and TG_(K) inanother is supported by the results of this analysis and by the genestructure and genomic organisation of the different transglutaminasegenes.

[0012] The inventors have therefore determined the structure of thehuman TGM5 gene, and its flanking sequences, and have mapped the gene tothe 15q15 region of chromosome 15. Further, the inventors havedetermined that the human TGM5 gene comprises 13 exons separated by 12introns spanning roughly 35 kb, and that the structure of the TGM5 geneis identical to that of EPB42 (band 4.2 protein), TGM2 (TG_(C)) and TGM3(TG_(E)) genes. Southern blot analysis has also shown that TGM5 is asingle copy gene in the haploid genome. The inventors developed a methodfor detection and identification of transglutaminase gene products basedon RT-PCR with degenerate primers and using this method have discoveredthe gene product of the TGM5 gene in keratinocytes (Aeschlimann et al(1998) J. Biol. Chem. 273, 3452-3460). Using this method, the inventorshave identified another new transglutaninase gene product in humanforeskin keratinocytes and in prostate cacrinoma tissue which has beendesignated TG_(Z) or transglustaminase type VII. A full-length cDNA forthis gene product was obtained by anchored PCR. Long range genomic PCRwas used comprising different combinations of primers designed from theflanking sequences of the TGM5-EPB42 gene sequence and the TG_(Z) cDNAsequence to explore whether the gene encoding TG_(Z) (TGM7) was presentin close proximity to the other two transglutaminase genes. This placedthe TGM7 gene approximately 9 kb upstream of the TGM5 gene anddemonstrated that the genes are arranged in tandem fashion (FIG. 5C).The inventors have therefore determined that the transglutaminase genes,TGM5 (TG_(X)), TGM7 (TG_(Z)) and EPB42 (band 4.2 protein) are positionedside by side within approximately 100 kb on chromosome 15. It has alsobeen found that the mouse homologues of these genes are similarlyarranged on mouse chromosome 2. Finally, the inventors have identifiedand determined the nucleotide and amino acid sequences as well as tissuedistribution for the novel transglutaminase gene products TG_(Z) andTG_(Y).

[0013] According to a first aspect of the invention there is provided anucleotide sequence comprising at least a portion of the nucleotidesequence of FIG. 6A, FIG. 6B, FIG. 10A or FIG. 10B; a nucleotidesequence which hybridise to the nucleotide sequence of FIG. 6A, FIG. 6B,FIG. 10A or FIG. 10B; a nucleotide sequence which is degenerate to thenucleotide sequence of FIG. 6A, FIG. 6B, FIG. 10A or FIG. 10B; all ofwhich nucleotide sequences encode a polypeptide having transglutaminaseactivity.

[0014] Preferably the nucleotide sequence consists of the nucleotidesequence of FIG. 6A, FIG. 6B, FIG. 10A or FIG. 10B.

[0015] The first aspect of the present invention also provides anucleotide sequence which hybridises under stringent conditions to thenucleotide sequences of FIG. 6A, FIG. 6B, FIG. 10A or FIG. 10B and whichencodes a polypeptide having transglutaminase activity. Preferably thenucleotide sequence has at least 80%, more preferably 90% sequencehomology to the nucleotide sequence shown in FIG. 6A, FIG. 6B, FIG. 10Aor FIG. 10B. Homology is preferably measured using the BLAST program.

[0016] The invention further provides a method of expressing apolypeptide comprising inserting a nucleotide sequence according to thefirst aspect of the present invention into a suitable host andexpressing that nucleotide sequence in order to express a polypeptidehaving transglutaminase activity.

[0017] The invention also provides a vector comprising a nucleotidesequence according to the first aspect of the present invention.

[0018] According to another aspect of the invention there is provided apolypeptide having an amino acid sequence comprising at least a portionof the amino acid sequence of FIG. 6A, FIG. 6B, FIG. 10A or FIG. 10B,wherein the polypeptide has transglutaminase activity.

[0019] The invention also provides a polypeptide sequence which is atleast 90% identical to the amino acid sequence of FIG. 6A, FIG. 6B, FIG.10A or FIG. 10B and which has transglutaminase activity. The amino acidsequence of the polypeptide having transglutaminase activity may differfrom the amino acid sequence given in FIG. 6A, FIG. 6B, FIG. 10A or FIG.10B by having the addition, deletion or substitution of some of theamino acid residues. Preferably the polypeptide of the present inventiononly differs by about 1 to 20, more preferably 1 to 10 amino acidresidues from the amino acid sequence given in FIG. 6A, FIG. 6B, FIG.10A or FIG. 10B.

[0020] The invention also provides a composition comprising thepolypeptide of the present invention for use in transamidation reactionson peptides and polypeptides.

[0021] The invention also provides a polypeptide comprising exons VIIthrough to exon X of the sequence shown in FIG. 6A or FIG. 6B. Theposition of the exons on the sequence shown in FIG. 6A or FIG. 6B can bedetermined from FIG. 8 where intron splice sites are marked with arrowheads.

[0022] According to a further aspect of the invention, there is provideda polypeptide comprising exons II through to exon IV or exons X throughto exon XII of the sequence show in FIG. 10A or FIG. 10B. As indicatedabove, the positions of the exons on the sequence shown in FIG. 10A orFIG. 10B can be determined from FIG. 8.

[0023] According to another aspect of the invention there is provided acomposition comprising the polypeptide according to the presentinvention for use in the cross-linking of proteins.

[0024] According to a further aspect of the invention there is provideda diagnostic method comprising detecting expression of the polypeptideaccording to the present invention in a subject or in cells derived froma subject.

[0025] The invention also provides an antibody directed against thepolypeptide according to the present invention. The antibody may be anyantibody molecule capable of specifically binding the polypeptideincluding polyclonal or monoclonal antibodies or antigen bindingfragments such as Fv, Fab, F(ab′)₂ fragments and single chain Fvfragments.

[0026] The invention further provides a method of gene therapycomprising correcting mutations in a non wild type nucleotide sequencecorresponding to the nucleotide sequence of FIG. 6A, FIG. 6B, FIG. 10Aor FIG. 10B. Such gene therapy methods can be performed by homologousrecombination techniques or by using ribozymes to correct small sequencemutation. Suitable techniques are well known to those skilled in theart.

[0027] In accordance with a further aspect of the invention there isprovided a method of diagnosis of autoimmune disease comprising taking asample from a subject and testing that sample for the presence of atransglutaminase encoded by the nucleotide sequence of FIG. 6A, FIG. 6B,FIG. 10A or FIG. 10B or portions thereof. Preferably thetransglutaminase is detected by using an antibody having affinity forthe transglutaminase.

[0028] The invention also provides a competitive protein binding assayfor the differential diagnosis of autoimmune diseases comprising thedetection of antibodies against the transglutaminase encoded by thenucleotide sequence of FIG. 6A, FIG. 6B or FIG. 10A or FIG. 10B, orportions thereof.

[0029] Preferably the protein binding assay comprises using exogenoustransglutaminase TG_(Z) or TG_(Y), or both, as a competitive antigen.

[0030] The invention will now be described with reference to theaccompanying FIGS. 1 to 11, in which:

[0031]FIG. 1 is a representation of the genomic organisation of thehuman TGM5 gene. The human TGM5 gene is represented with the exonsnumbered I to XIII indicated by solid boxes separated by the introns 1to 12. The sizes of the introns and exons are given in bp (base pairs).The 5′- and 3′-untranslated regions in exon 1 and XIII, respectively,are represented by hatched boxes with functional elements defining thetranscript indicated. Additional sequence elements found in the TGM5gene are indicated as follows: Alu, Alu 7SL derived retroposon; STS,sequence tagged site. Below the genomic map, a representation of thesequences present in the individual BAC clones is depicted.

[0032]FIG. 2 is a representation of the structure of the 5′ untranslatedregion of the human TGM5 gene and mapping of the transcriptional startsite. A. Primer extension analysis of poly (A⁺) RNA isolated fromprimary human keratinocyte prior to (lane 1) or after (lane 2) culturein suspension for 12h. Extension products were separated on denaturingpolyacrylamide gel alongside a Sanger dideoxynucleotide sequencingreaction of the appropriate genomic DNA fragment primed with the sameoligonucleotide. The transcriptional start site is indicated by thearrow. B. Nucleotide sequence of the proximal 5′ region of the TGM5gene, 5′ ends of mRNA from primary keratinocytes mapped by RACE areindicated by arrowheads. The major transcription start site identifiedby primer extension is highlighted with an asterisk (labelled +1).Consensus sequences for putative regulatory elements are underlined.

[0033]FIG. 3 is a representation of the structure of the 3′ untranslatedregion of the human TGM5 gene. 3′-flanking sequence is shown withsequences homologous to known consensus sequences for 3′ processing oftranscripts (AATAAA, CAYTG and YGTGTTYY) underlined. The terminationpoints of cDNA's isolated from human keratinocytes (Aeschlimann et al(1998) J. Biol Chem 273, 3453-3460) by 3′ RACE are indicated byarrowheads. A pair of inverted long repeat sequences is highlighted initalics.

[0034]FIG. 4 is a southern blot analysis of human genomic DNA hybridisedto genomic TG_(X) probes. Human genomic DNA was digested with BamHI,EcoRI, and HindIII restriction enzymes and hybridised with short³²P-labelled DNA fragments corresponding to intron 2 and flankingsequences of exon II and III (left panel) and exon X (right panel),respectively. The migration positions of the HindIII DNA size markers isindicated on the left.

[0035]FIG. 5 shows the chromosomal localisation of the human TGM5 geneby fluorescence in situ hybridisation. A. representative picture offluorescine-labelled genomic DNA of BAC-228(P20) (fluroescence, arrows)hybridised to metaphase spreads of human chromosome stained withpropidium iodide. B. An ideogram of banded chromosome 15 showing thelocalisation of the fluorescent signal on 13 chromosomes. C. Is aschematic map of the respective locus showing the organisation of thegenes encoding TG_(Z) (TGM5), band 4.2 protein (EPB4.2) and TG_(Z)(TGM7) as well as other genes [mitochondrial ATPase subunit Dpseudogene; D-type cyclin-interacting protein 1 (DIP); EST GenbankAA457639, AA457640)], L1 repetitive element and genetic markers.

[0036]FIG. 6 shows A. the nucleotide sequence and deduced amino acidsequence for human TG_(Z). B. an alternative nucleotide sequence anddeduced amino acid sequence for human TG_(Z). The initiation andtermination codons as well as polyadenylation signal (AATAAA) areunderlined.

[0037]FIG. 7 is a representation of the different tissue expressionpatterns for TG_(X), band 4.2 protein TG_(Y) and TG_(Z) in differentfetal and mature human tissues. Human tissue Northern dot blotnormalised for average expression of 9 different housekeeping genesprobed with a fragment corresponding to the C-terminal β-barrel domainsof TG_(X) (A), TG_(Z) (B), (C) TG_(Y) and band 4.2 (D). A diagramshowing the type of poly (A)⁺ RNA dotted onto the membrane is shown inpanel E.

[0038]FIG. 8 is a comparison of the structure of the different humantransglutaminase genes. A. is an alignment of the nine characterisedhuman gene products (TG_(X) TG_(Y), TG_(Z) (shown in FIG. 6A), TG_(C),TG_(E), band 4.2, factor XIII a-subunit, TG_(K), TG_(P),) is shown, withdashes indicating gaps inserted for optimal sequence alignment andunderlined residues representing amino acids conserved in at least fivegene products. The sequences are arranged to reflect thetransglutaminase domain structure, based on the crystal structure offactor XIII a-subunit. N-terminal propeptide domain (d1), β-sandwichdomain (d2), catalytic core domain (d3) and β-barrel domains 1 (d4) and2 (d5) (from top to bottom). Known intron splice sites are marked by^(t). B. is an alignment of the nine characterised human gene products(TG_(X), TG_(Y), TG_(Z) (shown in FIG. 6B), TG_(C), TG_(E), band 4.2,factor XIII a-subunit, TG_(K), TG_(P),) is shown, with dashes indicatinggaps inserted for optimal sequence alignment and underlined residuesrepresenting amino acids conserved in at least five gene products. Thesequences are arranged to reflect the transglutaminase domain structure,based on the crystal structure of factor XIII a-subunit. N-terminalpropeptide domain (d1), β-sandwich domain (d2), catalytic core domain(d3) and β-barrel domains 1 (d4) and 2 (d5) (from top to bottom). Knownintron splice sites are marked by arrowheads.

[0039]FIG. 9 is a phylogenetic tree of the transglutaminase gene familyand genomic organisation of the genes in man and in mouse. Sequenceswere aligned to maximise homology as shown in FIG. 8 except includingsequences from different species as available: h, human; m, mouse; r,rat. Note, the mouse sequence for TG_(X) ³ is at present incomplete andno information is available for the N-terminal domain. In panel A5, ahypothetical pedigree for the gene family is given that is consistentwith the data on the sequence relationship of the individual geneproducts (A) as well as with the data on the gene structure and genomicorganisation (B). Phylogenetic trees based on the amino acid sequencehomology of the gene products have been constructed using the NJ method(Saitou and Nei, 1987) of the PHYLIP software package for (1) theN-terminal β-sandwich domain (2) the catalytic core domain, (3) theC-terminal β-barrel domains, and (4) the entire gene products, (C).Shows the similarity of TG_(X) to the other transglutaminase geneproducts. The domain structure is based on the X-ray crystallographicstructure of the factor XIII a-subunit dimer and inferred on the othergene products based upon the sequence alignment shown in FIG. 8. Thenumbers reflect % sequence identity.

[0040]FIG. 10 shows the nucleotide sequence and deduced amino acidsequence of TG_(Y). A. Shows the nucleotide and deduced amino acidsequence for the long form of TG_(Y). B. Shows the nucleotide sequenceand deduced amino acid sequence for the short form of TG_(Y).

[0041]FIG. 11 is a schematic representation of the organisation of theidentified transglutaminase gene clusters in the human genome.

[0042] Isolation and Determination of the Structure of the Human TGM5Gene.

[0043] A unique insertion sequence of about 30 amino acids between thecatalytic core domain and β-barrel domain 1 found in TG_(X) was used asa template to design specific primers for the screening of a humangenomic library. The characterisation of several genes of thetransglutaminase gene family showed that the positions of the intronshas been highly conserved and a comparison of the TG_(X) sequence to thesequences of the other transglutaminases indicated that this uniquesequence is present within an exon, exon X (see FIG. 8, aa 460-503) inTG_(X). A PCR reaction from human genomic DNA using oligonucleotides P1and P2 that match sequences at either end of this unique segment yieldeda DNA fragment of expected size which was confirmed to be the correctproduct by sequencing (results not shown). Screening of a human genomicDNA BAC library by PCR using these oglionucleotides revealed twopositive clones, BAC-33(P5) and BAC-228(P20) that were subsequentlyshown by Southern blotting with different cDNA probes to containsequences spanning at least exon II to exon X of the TGM5 gene (resultsnot shown).

[0044] Restriction analysis further indicated that each of the BACclones contained substantially more than 50 kb of human genomic DNA.

[0045] The similarity in the gene structure of the differenttransglutaminase genes prompted us to approach the characterisation ofintrons by PCR amplification using oligonucleotide primers correspondingto the flanking exon sequences at the presumptive exon/intronboundaries. All intron/exon boundaries were sequenced from the PCRproducts obtained in at least two independent PCR reactions, whereapplicable from both BAC clones, to exclude mutations introduced by TaqDNA polymerase, and the results compared. When sequences of PCR productscomprising adjacent introns had no overlap, the intervening sequence(exon sequence) was determined by direct sequencing from isolated BACplasmid DNA to confirm the absence of additional introns. Similarly, the3′-untranslated region was obtained by step-wise extension of the knownsequence using direct sequencing of BAC plasmid DNA. Both BAC clonesterminated short of exon I and all attempts at isolating clones spanningexon I by screening of BAC and P1 libraries failed. Exon I and intron 1sequences were finally derived by nested PCR from human genomic DNAusing conditions optimised for long range genomic PCR.

[0046] We established that the TGM5 gene comprises approximately 35 kbof genomic DNA and contains 13 exons and 12 introns (FIG. 1). Allintron/exon splice sites conform to the known GT/AG donor/acceptor siterule and essentially to the consensus sequence proposed by Mount S. M(1982) Nucleic Acids Res. 10, 459-472. (Table I). A sequence homologousto the branch point consensus CTGAC (Keller E. B and Noon. W. A (1984)Proc. Natl. Acad. Sci. USA 81: 7417-7420) was found 24 to 44 nucleotidesupstream of the 3′ splice site in introns 1, 3-6, and 9-12. The size ofthe introns varied considerably, ranging from 106 bp to more than 6 kb(FIG. 1, Table II). The sequence obtained from the two different BACclones matched with the exception of a deletion spanning the sequencefrom intron 6 to intron 8 in BAC-33(P5) (FIG. 1).

[0047] Further, we also resequenced the entire coding sequence of TG_(X)and found 3 point mutations as compared to the previously reported cDNAsequence (Aeschlimann. et al (1998) J. Biol Chem 273 3452-3460) One ofthe nucleotide exchanges is silent, the other two result in an aminoacid exchange (Table III). The first two mutations were found in bothBAC clones, the third was only present in BAC-228(P20) due to thedeletion in the other BAC clone. These differences may result fromsequence polymorphisms in the human gene pool as there was no ambiguityof the cDNA-derived sequence in this position determined from multipleindependently amplified PCR products. However, the fact that a serineand alanine residue are changed into a proline and glycine residue thatconstitute the conserved amino acid in these positions in thetransglutaminase protein family (see FIG. 8, aa 67 and aa 352 in TG_(X))suggested that these may have been PCR-related mutations in the cDNAsequence. To clarify this issue, we have prepared cDNA from humanforeskin keratinocytes from different individuals, amplified full-lengthcDNA with high fidelity DNA polymerase, and sequenced the respectiveportions of the cloned cDNAs. The data confirmed that allelic variantsexist with differences in these positions (Table III).

[0048] The isolation and sequencing of cDNAs encoding TG_(X) andNorthern blotting with TG_(X) cDNA probes revealed expression of atleast two differentially spliced mRNA transcripts for TG_(X) in humankeratinocytes. Solving the gene structure confirmed the short form ofTG_(X) to be the result of alternative splicing of exon m as predicted.A third isolated cDNA that differed also at the exon III/exon IV splicejunction turned out to be the result of incomplete or absent splicingout of intron 3 as the sequence upstream of exon IV in the cDNA matchedwith the 3′ sequence of intron 3. Exon 3 encodes part of the N-terminalβ-barrel domain of TG_(X) and the absence of the sequence encoded byexon 3 is expected to result in major structural changes in at leastthis domain of the protein. Nevertheless, expression of TG_(X) in 293cells using the full-length cDNA resulted in synthesis of twopolypeptides with a molecular weight consistent with the predictedproducts from the alternatively spliced transcripts (results not shown).

[0049] Initially, 5′ RACE was used to determine the 5′ end of TG_(X)cDNAs. Transcripts starting 77,96 and 157 nucleotides upstream of theinitiator ATG were isolated in addition to the previously describedshorter transcript (FIG. 3B, arrowheads). All of these transcripts wererecovered repeatedly in independent experiments. Finally, primerextension experiments located the major transcription initiation siteused in keratinocytes 157 nucleotides upstream of the translation startcodon (FIG. 3). The proximal promoter region was analysed for potentialbinding sites of transcription factors using MatInspector (Genomatix,Munich Germany) and GCG (Genetics Computer Group, Inc., Madison, Wis.)software packages. No classical TATA-box sequence was found but a numberof other potential transcription factor binding sites could beidentified (FIG. 3b), suggesting that TG_(X) promoter is a TATA-lesspromoter. Interaction of C/BP (may bind to CAAT-box), nuclear factor I(NF1) and upstream stimulatory factor (USF) to form a core proximalpromoter has been demonstrated in a number of TATA-less genes. c-Myb isfound in TATA-less proximal promoters of genes involved in hematopoiesisand often interacts with Ets-factors, and these sites may be operativein the expression of TG_(X) in hematopoetic cells, e.g HEL cells AP1,Ets and SP1 elements are typically found in keratinocyte-specific genesand may be involved in transcriptional regulation in keratinocytes.Several AP1 sites are present within 2.5 kb of the upstream sequence andcould interact with the proximal AP1 factor for activation. SP1 sitesare properly positioned upstream of the start points of the shortertranscripts raising the possibility that these could also be functional,though to a lesser degree.

[0050] The last exon, exon XIII, contained a consensus polyadenylationsignal AATAAA-600 bp downstream of the termination codon (FIG. 3). Thisis in good agreement with the size of the mRNA (2.8 kb) encodingfull-length TG_(X) expressed in human keratinocytes as detected byNorthern blotting considering the length of the coding sequence (2160bp.). A CAYTG signal that binds to U4 snRNA which is identical for 4 outof 5 nucleotides is present in tandem in 3 copies 7 nucleotidesdownstream of the polyadenylation signal. A close match (YCTGTTYY) ofanother consensus sequence YGTGTTYY that is found in many eukaryotictranscripts and provides a signal for efficient 3′ processing is present46 nucleotides downstream of the polyadenylation signal. However, wehave previously reported that all cDNAs isolated by RT-PCR with anoligo(dT) oligonucleotide from human keratinocytes ended within 9 to 34nucleotides downstream of the pentanucleotide ATAAA at position 2169. Ithas been shown that this pentanucleoditde functions as a polyadenylationsignal and these shorter transcripts are selectively enhanced by PCRamplification because of the smaller size of the PCR product.

[0051] Chromosomal Localisation of the TGM5 Gene.

[0052] To address the genomic organisation and identify the chromosomallocalisation of the TGM5 gene or genes in the human genome, we performedSouthern blot analysis of human genomic DNA cut with BamHI, EcoRI andHindIII restriction enzymes using probes derived from intron 2 as wellas from the sequence encoded by exon X that is unique to TG_(X) (FIG.4). Bands of 4.5, 6.0, 10.5, 4.3, 9.3 and 2.6 kb were revealed with therespective probes. The simple pattern of restriction fragmentshybridising with the probes indicated that the haploid human genomecontains only one TGM5 gene.

[0053] The TGM5 gene was subsequently localised to chromosome 15 byfluorescent in situ hybridisation of human metaphase chromosome spreadsusing genomic DNA derived from either BAC clone as a probe (FIG. 5A). Acomparison of the probe signal to the DAPI banding pattern localised theTGM5 gene to the 15q15 region. The localisation was subsequently refinedby determining the distance of the fluorescent signal to the centromereas well as to either end of the chromosome on 13 copies of chromosome 15and expressing it as a fractional distance of the total length of thechromosome. These measurements placed the TGM5 gene close to the centreof the 15q15 region, i.e. to the 15q15.2 locus (FIG. 5B).

[0054] TGM5 is Part of a Cluster of Transglutaminase Genes.

[0055] The EPB42 gene has previously been assigned to locus 15q15.2 onchromosome 15. This raised the possibility that the EPB42 gene may bearranged in tandem with the TGM5 gene. Indeed, PCR with specific primersfor sequences derived from the 5′ and 3′ of the EPB42 gene yieldedproducts of appropriate size from both, BAC-33(P5) and BAC-228(P20)(results not shown) and sequencing confirmed the identity of the PCRproducts. Southern blotting of BAC plasmid DNA with cDNA probescomprising the 5′ or 3′ end of the EPB42 gene and subsequent comparisonof the pattern of labelled restriction fragments with that of the TGM5gene allowed us to map this locus in more detail. The EPB42 gene andTGM5 are arranged in the same orientation being spaced apart by ˜11 kb(FIG. 5C) approximately 30% of which was sequenced to characterise the3′ and 5′ flanking UTR of the TGM5 and EPB42 gene, respectively.

[0056] To analyse the relationship between the most closely homologousgenes of the transglutaminase gene family (TGM7, TGM5 and EPB42 on humanchromosome 15q15.2 and TGM2 and TGM3 on chromosome 20q11/12), we mappedthe respective mouse genes using radiation hybrid mapping. All genesmapped to the distal part of mouse chromosome 2. The genes for tgm7,tgm5, and epb42 showed a best fit location for the segment defined byD2Mit104 proximal and D2Mit305 (66.9cM) and an LOD of >20 to D2Ertd616e(69.0cM). This is in good agreement with the assigned locus 67.5cMdistal from the centromere, White, R. A., et al., (1992) Nat. Genet. 2,80-83. This tgm3 gene showed a best fit location for the segment definedby D2Mit447 proximal and D2Mit258 distal, with a highest LOD of 14.8 and12.2 to D2Mit258 (78.0cM) and D2Mit338 (73.9cM), respectively. The tgm2gene showed a best fit location for the segment defined by D2Mit139proximal (86.0cM) and D2Mit225 distal (91.0cM), with a highest LOD of17.0 to the anchor market D2Mit287, consistent with it's assigned locus89.0cM from the centromere, Nanda, N., et al., (1999). Arch. Biochem.Biophys. 366, 151-156.

[0057] We developed a method for detection and identification oftransglutaminase gene products based on RT-PCR with degenerate primersand using this method discovered the gene product of the TGM5 gene inkeratinocytes. Using this same method, we have identified another newtransglutaminase gene product in human foreskin keratinocytes and inprostate carcinoma tissue which we designated TG_(Z) or transglutaminasetype VII. A full-length cDNA for this gene product was obtained byanchored PCR (see below). We used long range genomic PCR with differentcombinations of primers designed from the flanking sequences of theTGM5-EPB42 gene segment and the TG_(z) cDNA sequence to explore whetherthe gene encoding TG_(z), TGM7, was present in close proximity to theother two transglutaminase genes. This placed the TGM7 geneapproximately 9 kb upstream of the TGM5 gene and demonstrated that thegenes are arranged in tandem fashion (FIG. 5C). The 5′ UTR of the TGM5gene was sequenced (FIG. 2). The genes encoding TG_(C), TG_(E), whichare more closely related to the TGM6, TGM7, and EPB42 genes than theother transglutaminase genes based on amino acid sequence comparison andsimilarity in gene structure (FIG. 9) have been mapped to humanchromosome 20q11 (Gentile V. et al (1994) Genomics 20, 295-297; Kim I.G. et al (1994) J. Invest. Dermatol 103, 137-142). The syntenic regionsof the 15q15 and 20q11 locus in the mouse are present in a shortsegment, 2F1-G, of chromosome 2, which puts all five transglutaminasegenes in proximity (FIG. 9B). The mouse homologue of band 4.2 proteinhas been mapped to this region of mouse chromosome 2 (White R. A. et al(1992) Nat. Genet 2, 80-83). We have isolated a BAC clone containing thegene encoding the mouse homologue of TG_(x) and shown that the tgm5 geneis located next to the epb42 gene in tandem fashion, similar to theorganisation in the human genome. Furthermore, this clone was shown tocontain the gene encoding the mouse homologue of TG_(z). Genomicsequences derived from this BAC clone and also cDNA sequences derivedfrom cDNA prepared from mouse uterus showed that the mouse and humangene products are 85% identical on the nucleotide level. To analyse therelationship between the transglutaminase genes in more detail, wecalculated the amino acid similarity (FIG. 9C) based on the sequencealignment shown in FIG. 8 and calculated evolutionary distances usingdifferent algorithms (FIG. 9A). All algorithms predicted a closerelationship between TG_(X) and TG_(E), and band 4.2 protein and TG_(C),raising the possibility that a single transglutaminase gene initiallylocally duplicated to generate a cluster of 3 genes, followed by aduplication of a larger segment of the chromosomal region, gave rise tothe organisation of the genes in mouse. In humans these chromosomalregions were apparently redistributed to two different chromosomes. Thishypothesis led us to spectulate on the existence of an additional geneon human chromosome 20q11. Careful analysis of the chromosomal sequencesof this locus derived by the human genome project revealed the presenceof a candidate gene, TGM6, located approximately 45 kb downstream of theTGM3 gene consistent with our hypothesis (FIG. 11). To confirm that thisis in fact a functional gene and not a pseudogene, we screened a largenumber of cell lines for expression of a respective gene product by PCR.A corresponding gene product, TG_(Y), or transglutaminase type VI couldbe identified in a small cell lung carcinoma cell line and a full-lengthcDNA was subsequently derived by anchored PCR.

[0058] Determination of cDNA and Amino Acid Sequences of TGM6 (TG_(Y))and TGM7 (TG_(Z)) Gene Products

[0059] A full-length cDNA sequence for TG, was obtained by anchored PCRusing oligo(dT)-Not I primed cDNA prepared from human foreskinkeratinocytes, prostate carcinoma tissue and human carcinoma cell linePC3, essentially following the strategy previously described(Aeschlimann et al (1998) J. Biol Chem 273: 3245-3460). Theoligo(dT)-Not I primer was used as the anchoring primer to obtain the 3′end of the cDNA. 5′ RACE was used to determine the 5′ end of the cDNA.The obtained sequence information (FIG. 6) contained an open readingframe 2130 nucleotides and a polyadenylation signal (AATAAA) 158nucleotides downstream of the termination codon (TGA). The deducedprotein consists of 710 amino acids. The cDNA and amino acid sequence inFIG. 6A was first determined and the deduced protein has a calculatedmolecular mass of 79,908 Da and an isoelectric point of 6.7. The cDNAand amino acid sequence in FIG. 6B was then determined. This sequencediffers by a few nucleotides and amino acids from the sequence given inFIG. 6A. The protein deduced from the sequence given in FIG. 6B has acalculated molecular mass of 80,065 and an isoelectric point of 6.6. Anumber of aberrantly spliced gene products were isolated which lackedpart of exon IX (5′end) or-retained the whole or part of intron 11.These products are unlikely to be of physiological significance but maypoint out that splicing of certain introns in this gene is a difficultand inefficient process.

[0060] A full-length cDNA sequence for TG_(Y) was obtained by PCR usingoligo(dT) primed cDNA prepared from the lung small cell carcinoma cellline H69, and using sequence specific primers based on the presumptivetranscribed genomic sequence. 5′ RACE was used to determine the 5′ endof the cDNA. The obtained sequence information for the long form ofTG_(Y) (FIG. 10A) contained an open reading frame of 2109 nucleotides.The deduced protein for the long form of TG_(Y) consists of 708 aminoacids and has a calculated molecular mass of 79,466 Da and anisoelectric point of 6.9. A shorter transcript was also isolated whichapparently resulted from alternative splicing of the sequence encoded byexon XII. The absence of exon XII results in a frame shift and therebyin premature termination within exon XIII. The obtained sequenceinformation for the short form of TG_(Y) (FIG. 10B) contained an openreading frame of 1878 nucleotides. The deduced protein for the shortform of TG_(Y) consists of 626 amino acids and has a calculatedmolecular mass of 70,671 Da and an isoelectric point of 7.6. Thesequence alterations due to the splicing result in a short protein whichterminates just after the first C-terminal β-barrel domain. The β-barreldomains have been implicated in the regulation of enzyme-substrateinteraction, and the lack of the second C-terminal β-barrel domain (seeFIG. 8, d5) is likely to be of biological significance.

[0061] The catalytic mechanism of transglutaminases has been solvedbased on biochemical data available for several transglutaminases andthe X-ray crystallographic structure of the factor XIII a-subunit dimer.The reaction center is formed by the core domain and involveshydrogen-bonding of the active site Cys to a His and Asp residue to forma catalytic triad reminiscent of the Cys-His-Asn triad found in thepapain family of cysteine proteases. The residues comprising thecatalytic triad are conserved in TG_(Y) (Cys276, His335, Asp358) andTG_(Z) (Cys227, His336, Asp359) (FIG. 8) and the core domain shows ahigh level of conservation as indicated by a sequence identity of about50% between these gene products and the other transglutaminases FIG. 9).A Tyr residue in barrel 1 domain of the a subunit of factor XIII ishydrogen-bonded to the active site Cys residue and it has been suggestedthat the glutamine substrate attacks from the direction of this bond toinitiate the reaction based on analogy to the cysteine proteases. InTG_(Y), the Tyr residue is conserved (Tyr 540) while in TG_(Z) the Tyrresidue has been replaced by His538 similar to TG_(X) (FIG. 8). This isexpected to be a conservative change which is supported by our datademonstrating that recombinant TG_(X) from 293 cells hastransglutaminase activity. Crystallization experiments with factor XIIIafurther indicated that 4 residues are involved in binding of Ca²⁺-ion,including the main chain carbonyl of Ala457 and the side chain carboxylgroups of Asp438, Glu485, and Glu490. All three acidic residues areconserved in TG_(Y) and in TG_(Z) (FIG. 8). None of the residuescritical to enzyme function are affected by the alternative splicing ofTG_(Y). Based on the preservation of critical residues for enzymefunction and domain folding and the extensive overall similarity of theTG_(Y) isoforms and TG_(Z) to the other members of the transglutaminaseprotein family, it can be predicted that the characterized cDNAs areencoding active transglutaminases.

[0062] Tissue Expression Patterns for TG_(X), TG_(Y) and TG_(Z).

[0063] We have previously shown that TG_(X) is expressed in a number ofdifferent cell types (Aeschlimann et al (1998) J. Biol. Chem. 273,3452-3460). To obtain a more complete picture on the expression ofTG_(X) and the novel gene products, we performed a dot blot Northernblot analysis of more than 50 adult and fetal human tissues. Band 4.2protein was expressed at high level in bone marrow and fetal spleen andliver, consistent with its role in hematopoietic cells, and virtuallyundetectable in all other tissues. In contrast, TG_(X), TG_(Y) andTG_(Z) showed widespread expression at low level, with highest levels ofTG_(X), TG_(Y) and TG_(Z) mRNA present in the female reproductivesystem, in the central nervous system, and in testis, respectively (FIG.7).

[0064] RT-PCR analysis on human cell lines and tissues shows that TG_(Z)is expressed in osteosarcoma cells (MG-63), dermal fibroblasts (TJ6F,HCA2), erythroleukemia cells (HEL), in primary keratinocytes, mammaryepithelium carcinoma cells (CF7), HELA cells, skin, brain, heart,kidney, lung, pancreas, placenta, skeletal muscle, fetal liver, prostateand in prostate carcinoma tissue. A similar analysis for TG_(Y) revealedexpression only in a lung small cell carcinoma cell line (H69) andextremely low levels of expression in tissues.

[0065] In conclusion, TG_(Z) is expressed widely in cells and tissuesand expression levels are not apparently affected by cellulardifferentiation, (i.e keratinocyte differentiation or fibroblastsenescence). TG_(Y) expression, on the other hand, was very restrictedand expression was only found in H69 cell line. This cell line hascharacteristics of neuronal cells such as the expression ofneuron-specific enolase and brain isozyme of creatine kinase whichtogether with widespread expression in tissues of the nervous systemsuggests that TG_(Y) expression may be specific to neuronal cells.Transglutaminase action has been implicated in the formation of aberrantprotein complexes in the central nervous system leading to nerve celldegeneration, e.g in Alzheimers and Huntington's disease. Based on itsexpression pattern, TG_(Y) is a logical candidate to bring about theunderlying transglutaminase-related pathological changes.

[0066] Reagents

[0067] Oligonucleotides were from Oligos. Etc. Inc. (Wilsonville, Oreg.)or life technologies and restriction enzymes from Promega Corp.(Madison, Wis.).

[0068] Genomic Library Screening

[0069] A human BAC library established in a F-factor-based vector,pBeloBAC 11, and maintained in E. coli DH10B was screened by PCR (GenomeSystems, Inc., St. Louis, Mo.). A 147 bp DNA fragment unique to TG_(X)was amplified from 100 ng of genomic DNA in 100 μl of 10 mM Tris/HCl, pH8.3, 50 mM KCl containing 2 mM MgCl₂, 0.2 mM dNTPs using 2.5 units ofTag DNA polymerase (Fisher Scientific Corp. Pittsburgh, Pa.) and 50 pmolof upstream primer P1,5′-CCACATGTTGCAGAAGCTGAAGGCTAGAAGC and downstreamprimer P2,5′-CCACATGTCCACATCACTGGGTCGAAGGGAAGG. PCR cycles were 45 secat 94° C. (denaturation), 2 min at 60° C. (annealing), and 3 min at 72°C. (elongation) for a total of 37 cycles, with the first cyclecontaining an extended denaturation period (6 min) during which thepolymerase was added (hot start), and the last cycle contained anextended elongation period (10 min). Two positive clones wereidentified, BAC-33(P5) and BAC-228(P20) (Genome Systems), and theiridentity verified by Southern blotting. Plasmid DNA was prepared using astandard alkaline lysis protocol. 2 μg plasmid DNA was restricted withBamHI, EcoRI, and SpeI and probed with a ³²P-labelled-500 bp NcoI/BspHIand ˜600 bp BspHI/NdeI cDNA fragment of TG_(X), respectively, asdescribed below.

[0070] Amplification of TGM5 Intron Sequences

[0071] PCRs were carried out with 2.5 units of Taq DNA polymerase(Fisher Scientific) and 100-200 ng of plasmid DNA from BAC clones in 100μl of 10 mM Tris/HCl, pH 8.3 50 mM KCl containing 2 mM MgCl₂, 0.2 mMdNTPs and 50 pmol of the desired oligonucleotide primers. The PCR cycleswere 45 sec at 94° C. (denaturation), 1 min at 60° C. (annealing), and 5min at 72° C. (elongation). A total of 32 cycles were carried out, withthe first cycle comprising an extended denaturation period (6 min)during which the polymerase was added (hot start) and the last cyclecomprised an extended elongation period (10 min). The followingoligonucleotides were used as upstream and downstream primers,respectively, in the individual reactions:

[0072] intron 2,5′-GGACCACCTGCTTGTTCGCCGGGG,5′-AGGGGCTGGGGCTGTGATGGCGTG;

[0073] intron 3,5′-ACCTCITGAAAATCCACATCGACTCCT,5′-CAGTTCTTGCTGCCTTGGTAGATGAAGCC;

[0074] intron 4,5′-GACAGTGAACCCCAGAGGCAGGAG,5′-TCTGTGGCTGGGTCAGTCTGGAAGTGCA (P3);

[0075] intron 5,5′-GCCTGCACTTCCAGACTGACCCAGCCACA,5′-TCCAGTITCCATTGAGCACCCCA;

[0076] intron 6,5′-TGCTGGGTCTTTGCTGCCGTCATGTGC,5′-TCCTTTCTTTATTCCCCAAAATCCTGCC;

[0077] intron 7,5′-TAGATGAGTATTATGACAACACAGGCAGG,5′-GCGTCCAGCACCTGCCAGCCTCC;

[0078] intron 8,5′-TGAGTGCTGGATGGCCCGGAAGG, 5′-CCCGCTCGTCACTCTGGATGCTC;

[0079] intron 9,5′-TTCACCAGGACACGAGTTCTGTTGGCA, P2 (see above);

[0080] intron 10, P1 (see above), 5′-TCAGGACTGCTTTTCTCTTCACCC;

[0081] intron 11, 5′-ACCCCTGCAAAATCTCCTATTCCC,5′-AATATCACCTGTATGGAGAGTGGCTGG;

[0082] intron 12,5′-TTGAGGACTGTGTGCTGACTGTGGM5′-AATGATGCTTGCTTGGTGTTGGGG.

[0083] PCR's were carried out with 1.25 units of Pfu Turbo DNApolymerase (Stratagene) and 260 ng genomic DNA in a total of 100 μl ofsupplied reaction buffer supplemented with 0.2 mM dNTPs, 2 μl DMSO and50 pmol primers. The PCR cycles were 45 sec at 94° C. (denaturation), 1min at 68° C. (annealing), and 2 min at 72° C. (elongation). A total of37 cycles were made, with the first cycle containing an extendeddenaturation period (6 min) during which the polymerase was added (hotstart), and the last cycle containing an extended elongation period (10min).

[0084] Rapid Amplification of 5′-mRNA End

[0085] A modified RACE protocol was used to determine the transcriptionstart site and obtain additional sequence information of exon I. Doublestranded cDNA was prepared from poly(A⁺) RNA of cultured normal humankeratinocytes (Aeschilmann et al (1998) J. Biol. Chem. 273, 3452-3460)with the Copy Kit (Invitrogen, San Diego, Calif.). The cDNA was purifiedfrom nucleotides using the GlassMax DNA Isolation Kit (LifeTechnologies, Inc.) and tailed in the presence of 200 μM dCTP with 10units of terminal deoxynucleotidyl transferase (Promega) for 30 min at37° C. to anchor the PCR at the 5′-end. The PCR reaction was anchored byperforming a total of 5 cycles of one-sided PCR at a lower annealingtemperature (37° C.) with the abridged anchor primer (Life Technologies,Inc.) only. Following transfer of 25% of this reaction at 94° C. to anew tube containing abridged anchor primer and TG_(X)-specific primer P3(see above), the first round of amplification was carried out for atotal of 37 cycles under the conditions described above except forannealing which was carried out at 55° C. Nested PCR was done with theuniversal amplification primer (Life Technologies, Inc.) andTG_(X)-specific primer P4, 5′-TGAAGTACAGGGTGAGGTTGAAGG, as describedabove (annealing at 60° C.) using 1.0 μl from the first round PCR.

[0086] Primer Extension Analysis

[0087] Oligonucleotide P55′-CATGGTAGCTGCCTCCGGTTCCTG containing a5′-infrared label (IRD 800) was purchased from MWG Biotech (Ebesberg,Germany). Primer P5 (5.3 pmol) was hybridised to 1 μg of poly (A⁺) RNAfrom primary keratinocytes (Aeschlimann et al (1998) J. Biol. Chem. 273,3452-3460) and reverse transcription performed with 200 units ofSuperscript II RNAse H reverse transcriptase (Life Technologies) in atotal of 20 μl for 90 min at 42° C. according to the manufacturer'sinstructions. Enzyme was heat inactivated and primer extension productsextracted with phenol chloroform, precipitated with ethanol, and thenanalysed on a 4.5% denaturing polyacrylamide gel adjacent todideoxynucleotide chain termination sequencing reactions (ThermoSequenase Cycle Sequencing Kit; Amersham) derived from a double-strandedgenomic DNA fragment using the same primer.

[0088] DNA Preparation and Sequencing

[0089] Plasmid DNA from BAC clones was further purified for directsequencing by digestion with 200 μg/ml of RNase A (Sigma, St. Louis,Mo.) for 1h at 37° C. and by subsequent micro-dialysis using Spectra/Por2 membranes (Spectrum Medical Industries, Inc. Laguana Hills, Calif.).PCR produts were gel purified using the QIA quick Gel Extraction Kit(Qiagen, Inc. Chatsworth, Calif.) for sequencing. Cycle sequencing wasperformed by the dideoxy chain termination method using the CyclistExo-Pfu DNA Sequencing Kit (Stratagene, LaJolla, Calif.) and pre-cast 6%polyacrylamide gels with the CastAway Sequencing System (Stratagene) orusing the dRhodamine Terminator Cycle Sequencing Ready Reaction Kit (PEBiosystems) and an ABI 310 automated sequencer.

[0090] Southern Blotting

[0091] 18%1 g human genomic DNA was digested with BamHI, EcoRI, andHindIII restriction enzymes, separated in a 0.8% agarose gel andtransferred to a Zeta-probe membrane (Bio-Rad, Labs. Hercules, Calif.).The gel was calibrated using the Lambda DNA/HindIII markers (Promega).³P-labelled probes were prepared by random prime labelling using theMultiprime DNA Labelling System (Amersham, Int. Amersham, UK) and PCRproducts corresponding to intron 2, intron 12, and exon X (see above) asDNA templates. Probes were hybridised to the blot overnight at 65° C. in500 mM NaH₂PO₄, pH 7.5, containing 1 mM EDTA and 7% SDS. The membranewas washed at 65° C. to a final stringency of 40 mM NaH₂PO₄, pH 7.5, 1mM EDTA, and 1% SDS, and the result developed by exposure of themembrane to BioMax MR film (Eastman Kodak, Rochester, N.Y.).

[0092] Chromosomal Localisation

[0093] Human peripheral blood lymphocytes were used to prepare metaphasechromosome spreads (Bebbington C. R. and Hentschel, C. C. G. (1987) inDNA cloning (Volume III) 184-188, IRL Press, Oxford UK). Cells werecultured in PB-Max Karyotyping medium (Gibco, BRL, Gaithersburg, Md.)for 72h, and synchronised by culture in the presence of 10⁻⁷Mamethophterin (Fluka) for another 24h. Cells were released from themitotic block by extensive washing and subsequent culture in the abovemedium containing 10⁻⁵M thymidine for 5h. Cells were subsequentlyarrested in metaphase by addition of colcemid to a final concentrationof 0.1 μg/ml (Gibco BRL). Harvested cells were incubated in 0.075M KCPfor 25 min at 37° C., fixed in methanol/acetic acid (3:1) solution, andchromosome spreads prepared by dropping the cells onto the glass slides.After air drying, chromosomes were treated with 1001 g/ml of RNase A in2×SSC for 1 h at 37° C., denatured in 70% (v/v) formamide in 2×SSC for 3min at 75° C., and dehydrated in a graded ethanol series. DNA probeswere prepared by random prime labelling of plasmid DNA of BAC-33(P5) andBAC-228(P20) with fluorescine-conjugated dUTP using the Prime-It FluorFluorescence Labelling Kit (Stratagene). Probes were denatured at 75° C.for 10 min in hybridisation buffer consisting of 50% formamide (v/v) and10% dextran sulphate (w/v) in 4×SSC and prehybridised at 42° C. for 20min to 0.2 μg/l human competitor DNA (Stratagene) to block repetitiveDNA sequences. Probes were subsequently hybridised to the chromosomespreads at 37° C. overnight, followed by washing to a final stringencyof 0.1×SSC at 60° C. Spreads were mounted in phosphate-buffered glycerolcontaining 200 ng/ml propidium iodide to counterstain chromosomes.Slides were examined by epifluorescence microscopy using a 100×objective and images captured with a DC-330 CCD camera (DAGE-MTI, Inc.Michigan City, 1N) using a LG-3 frame grabber board (Scion Corp.Frederick, Md.) in a McIntosh 8500 workstation and a modified version ofthe NIH image 1.6 software (Scion Corp.). Images representingfluorescine-labelling and propidium iodide staining of the same fieldwere superimposed using Adobe Photoshop 3.0 (Adobe Systems, Inc.Mountain View, Calif.) to map the gene to a chromosomal region.

[0094] Cloning of Novel Transglutaminase Gene Products by Anchored PCR

[0095] For cloning of TG_(Y), poly(A)+ RNA was prepared from about106H69 cells (American Type Culture Collection, Rockville, Md.) byoligo(dT)-cellulose column chromatography using the Micro-Fast Track Kit(Invitrogen, San Diego, Calif.) and recovered in 20 μl 10 mM Tris/HCl,pH 7.5. The poly(A)+ RNA (5.0 μl) was reverse transcribed into DNA in atotal volume of 20 μl using the cDNA Cycle Kit (Invitrogen) with 1.0 μloligo(dT) primer (0.2 μg/μl). Overlapping fragments of TG_(Y) wereamplified by PCR using oligonucleotides 5′-ATCAGAGTCACCAAGGTGGAC,5′-AGAAACACATCGTCCTCTGCACACC (P6), 5′-CAGGCTTTCCTCTCACCGCAAACAC,5′-CGTACTTGACTGGCTTGTACCTGCC, 5′-TCTACGTCACCAGGGTCATCAGTGC,5′-GCCTGTTCACCGCCTTGCTGT, 5′-CATCACTGACCTCTACAAGTATCC,5′-ACGGCGTGGGATTCATGCAGG, 5′-CATCCTCTATACCCGCAAGCC, and5′-AGGTTGAGGCAGGATTAACTGAGGCCTC. PCRs were carried out with 1.25 unitsof AmpliTaq Gold DNA polymerase (PE Biosystems) and 2.0 μl cDNA in atotal of 50 μl of supplied reaction buffer supplemented with 2 mM MgCl₂,0.2 mM dNTPs and 25 pmol of the appropriate gene-specific primers. Atotal of 40 PCR cycles were made, with an elevated annealing temperatureof 65° C. for the initial 5 cycles and an annealing temperature of 60°C. for the remaining cycles. The 5′-end of the cDNA was isolated by5′-RACE as described above with the exception of using the gene-specificoligonucleotides P6,5′-GATGTCTGGAACACAGCTTTGG, and5′-TCACAGTCCAGGGCTCTGCTCAG. The PCR-products were either directlysequenced or when desired, cloned by taking advantage of the 3′A-overhangs generated by Taq DNA polymerase using the OriginalTA-Cloning Kit (Invitrogen).

[0096] For cloning of TG_(Z), we used a series of degenerate andgene-specific oligonucleotides to isolate overlapping DNA fragments,essentially following our previously described strategy. TG_(Z)-specificoligonucleotide primers were 5′-CAACCTTGCGGCTTGAGTCTGTCG,5′-CAGCAGCTCTGACGGCTTGGGTC (P7), 5′-ATCACCTTTGTGGCTGAGACCG,5′-CAAGGGTTAAAAAGTAGGATGAAAGTTC, 5′-CACAGTGTGACTTACCCGCTG,5′-CATACACCACGTCGTTCCGCTG, 5′-CTTAAAGAACCCGGCCAAAGACTG,5′-CGATGGTCAAGTTCCTATCCAXGTTG, 5′-TGTTGTTTCCAATTTCCGTTCCGC,5′-TCTGGCACCCTCTGGATACGCAG, 5′-CTTAGGGATCAGCCAGCGCAGC,5′-GCGGATGAACCTGGACTTTGG, 5′-GGGTGACATGGACTCTCAGCG,5′-TGGGCAAGGCGCTGAGAGTCCATG, 5′-GCTGGAGGGCGGGTCTCAGGGAGC, and5′-AGGACAGAGGTGGAGCCAAGACGACATAGCC. Preparation of cDNA from humanforeskin keratinocytes and prostate carcinoma tissue has been describedpreviously. The PCRs were performed under the conditions described aboveor for PCR with degenerate primers as described previously. Nested PCRswere done by replacing the cDNA with 1.0 μl from the first PCR reaction.The 5′-end of the cDNA was isolated by 5′-RACE as described above withthe exception of using the gene-specific oligonucleotidesP7,5′-TGAAGCTCAGCCGGAGGTAGAAG, and 5′-GACAGACTCAAGCCGCAAGGTTG.

[0097] Northern Hybridization

[0098] A human RNA Master Blot containing poly(A)⁺ mRNA of 50 differenttissues was obtained from Clonetech Laboratories, Inc (Palo Alto,Calif.). ³²P-labeled probes were prepared by random prime labelling ofDNA fragments of the different transglutaminase gene products using theMultiprime DNA Labelling System (Amersham, Int., Amersham, UK). DNAfragments of 500-700 bp compromising the 3′-end of TG_(X), TG_(Z), andband 4.2 protein, were generated by restriction with Pst I and Acc L NcoI and Not I (exon XII and XIII), and Xho L respectively. The cDNAencoding human band 4.2 protein (Korsgren et al. 1990) was kindlyprovided by Dr Carl M. Cohen, Boston, Mass. A 220 bp ³²P-labeledfragment of TG_(Y) was generated by PCR using oligonucleotides5′-CAGCCTCAGTCACCGCCATCCGC and 5′-GATACTTGTAGAGGTCAGTGATG. Hybridizationwas performed under the conditions recommended by the manufacturer. Thelabeled membrane was exposed to BioMax MR film (Eastman Kodak) and filmsdeveloped after 15 to 24 hr for first exposure and 3 to 5 days forsecond exposure.

[0099] Amplification of TG_(Y) and TG_(Z) From Different Tissues

[0100] cDNA from various cell lines and human tissue was prepared aspreviously described. A panel of cDNAs from human tissue (MultipleTissue cDNA Panel I) were also obtained from Clonetech Laboratories. A365 or 287 bp fragment of TG_(Z) was amplified by PCR usingoligonucleotides 5′-TGGGCAAGGCGCTGAGAGTCCATG and5′-GCTGGAGGGCGGGTCTCAGGGAGC or 5′-AGGACAGAGGTGGAGCCAAGACGACATAGCC,respectively, with an annealing temperature of 60° C. A 218 or 170 bpfragment of TG_(Y) was amplified by PCR using oligonucleotides5′-CAGCCTCAGTCACCGCCATCCGC and 5′-GATACTTGTAGAGGTCAGTGATG or5′-GTGAAGGACTGTGCGCTGATG and 5′-CGGGAAGTGAGGGCTTACAAG, respectively, andidentical conditions as above.

[0101] Mapping of Transglutaminase Genes in Mouse Genome

[0102] The 100 radiation hybrid (RH) clones of the T31 mouse/hamster RHpanel (McCarthy et al., (1997), Genome Res., 7, 1153-1161) (ResearchGenetics, Huntsville, Ala.) were screened by PCR. A 139 bp fragment ofthe tgm5 gene was amplified with primers5′-TGAGGACTGTGTGCTGACCTTG (f)and 5′-TCCTGTGTCTGGCCTAGGG (r), a 149 bp fragment of the epb42 gene withprimers 5′-CAGGAGGAGTAAGGGGAATTGG (f) and 5′-TGCAGGCTACTGGAATCCACG (r),a 400 bp fragment of tgm7 with primers 5′-GGGAGTGGCCTCATCAATGG (f) and5′-CCTTGACCTCACTGCTGCTGA (r), a −600 bp fragment with tgm3 with primers5′-TCGGTGGCAGCCTCAAGATTG (f) and 5′-AGACATCAATGGGCAGGCATGG (r), and 655bp and 232 bp fragments of tgm2 with primers 5′-TTGGGGAGCTGGAGAGCAAC (f)and 5′-ATCCAGGACTCCACCCAGCA (r) and primers5′-(GCGGCCGCTAGT)CCACATTGCAGGGCTCCTGACT (f) and5′-GCTAGCCTGTGCTCACCATGAGG (r), respectively. PCRs were carried out in aGeneAmp 9600 thermacycler with 0.035 units/μl AmpliTaq Gold polymerasein standard reaction buffer containing 2 mM MgCl₂, 0.2 mM dNTPs, 0.4 μMof each primer and 2.5 ng/μl genomic DNA in a total reaction volume of25 μl. PCR conditions were: polymerase activation for 10 min at 95° C.,annealing at 60° C. for 45 sec, extension at 72° C. for 1 min anddenaturation at 94° C. for 30 sec for 35 cycles with a final extensionof 3.5 min at 72° C. PCR reactions were analyzed by agarose gelelectrophoresis using 1% or 1.5% gels. The hybrid cell panel wasanalyzed at least twice in each case to exclude PCT related errors. Thedata was submitted to the Jackson Laboratory Radiation Hybrid Databasefor analysis and mapped relative to known genomic markers(http://www.jax.org/resources/documents/cmdata.rhmap). TABLE I Splicedonor and acceptor sequences in the human TGM5 gene. Residues consistentwith the splice site consensus sequence (MAG/GTRAG and YAG/G) areunderlined. Intron number Donor sequence Acceptor sequence 1       M  A  Q                               G  L  E  V  AGCTACCATGGCCCAAGgtagggaaagcccctgtggccactggagttttttgtctaaccctggctgccccattgcagGGCTAGAAGTGGCC 2  F  V  V  E  T                              G  P  L  P  DTTCGTGGTTGAAACTGgtaagaaccccagctggctcacaggggctgtggagggcctcagctctacttccctcctagGACCGCTGCCAGAC 3  N  P  W  C  P                              E  D  A  V  YAATCCCTGGTGCCCAGgtaaggctgggtgcccaggcggtgcctccttgcttcgtgccctcccactctggttcctagAGATGCTGTCTAC 4  W  N  Y  G  Q                               F  E  D  K  ITGGAACTATGGACAGgtgagtctcagccctgcttatggcccatcctgccttccctctctgcctctccccccgaagTTTGAAGACAAAATC 5  V  V  C  A  M                               I  N  S  N  DGTGGTGTGTGCCATGgtgaggtccctggcgtgcccggggaggaggctcacacttctctatatggcttctcttcagATCAACAGCAATGAT 6  A  V  M  C  T                              V  M  R  C  LGCCGTCATGTGCACAGgtaggaggtagaaaggacctcacaaaaaggacaggtgatttttttgtgccctttttgcagTGATGAGGTGTCTG 7  K  D  T  I  W                                N  F  H  V  WAAGGATACTATCTGgtgagaaacaacctctcaacctatttctagcaacgtctcccttggctctgtttgatacagGAACTTCCATGTCTGG 8  Q  E  M  S  N                              G  V  Y  C  CCAGGAGATGAGCAACGgtgaggctctccagaagaaaggcaggccccgcccaccgaggctcccctgttctccttcagGCGTCTACTGCTGT 9  Y  K  Y  E  E                              G  S  L  Q  ETACAAGTATGAAGAAGgttagtaagcaagccagccctactcagagccagctggtgctgtgctctccccaacttcagGATCCCTCCAGGAG 10  L  S  P  K  E                              A  K  T  Y  PCTCTCTCCTAAAGAAGgtacgcatgtgcacagtttgtgtacgcagatctcaccccatccttgtgttctttctttagCAAAGACCTACCCC 11  S  I  T  I  N                               V  L  G  A  AAGCATCACGATTAATgtaggcaggagtcctgcaaatggcttgtggtaattctccttcccctcctggtctgtttagGTTCTAGGAGCAGCC 12  Q  Q  K  V  F                                L  G  V  L  KCAGCAGAAAGTCTTgtaagtgctgcaagtgctcagccttctcctttttctgacatgctccattctctgttgcagCCTTGGAGTCCTCAAA

[0103] TABLE II Intron sizes and splice types in the human TGM5 gene.Sizes of introns are estimated to be within about a 100 bp unlessindicated to be sequenced entirely. Intron number Splice type SizeMethod 1 1 6,300 bp PCR 2 1 102 bp Sequencing 3 1 3,300 bp PCR 4 0 2,900bp PCR 5 0 600 bp PCR 6 1 ˜11,800 bp PCR and Restriction Analysis 7 21,600 bp PCR 8 1 106 bp Sequencing 9 1 2,900 bp PCR 10 1 545 bpSequencing 11 0 1100 bp PCR 12 2 209 bp Sequencing

[0104] TABLE III Apparent polymorphisms in the cDNA and genomic DNAsequences for TG_(x). The positions with nucleotide and amino acidvariations are underlined. Residue cDNA (a) Gene cDNA (b) 67 S TCA P CCAP CCA 220 Y TAC Y TAT Y TAC 352 A GCA G GGA A GCA

[0105]

1 144 1 31 DNA Homo sapiens 1 ccacatgttg cagaagctga aggctagaag c 31 2 33DNA Homo sapiens 2 ccacatgtcc acatcactgg gtcgaaggga agg 33 3 24 DNA Homosapiens 3 ggaccacctg cttgttcgcc gggg 24 4 24 DNA Homo sapiens 4aggggctggg gctgtgatgg cgtg 24 5 27 DNA Homo sapiens 5 acctcttgaaaatccacatc gactcct 27 6 29 DNA Homo sapiens 6 cagttcttgc tgccttggtagatgaagcc 29 7 24 DNA Homo sapiens 7 gacagtgaac cccagaggca ggag 24 8 28DNA Homo sapiens 8 tctgtggctg ggtcagtctg gaagtgca 28 9 29 DNA Homosapiens 9 gcctgcactt ccagactgac ccagccaca 29 10 23 DNA Homo sapiens 10tccagtttcc attgagcacc cca 23 11 27 DNA Homo sapiens 11 tgctgggtctttgctgccgt catgtgc 27 12 29 DNA Homo sapiens 12 tccttcttct tattccccaaaatcctgcc 29 13 29 DNA Homo sapiens 13 tagatgagta ttatgacaac acaggcagg29 14 23 DNA Homo sapiens 14 gcgtccagca cctgccagcc tcc 23 15 23 DNA Homosapiens 15 tgagtgctgg atggcccgga agg 23 16 23 DNA Homo sapiens 16cccgctcgtc actctggatg ctc 23 17 27 DNA Homo sapiens 17 ttcaccaggacacgagttct gttggca 27 18 24 DNA Homo sapiens 18 tcaggactgc ttttctcttcaccc 24 19 24 DNA Homo sapiens 19 acccctgcaa aatctcctat tccc 24 20 27DNA Homo sapiens 20 aatatcacct gtatggagag tggctgg 27 21 25 DNA Homosapiens 21 ttgaggactg tgtgctgact gtggm 25 22 24 DNA Homo sapiens 22aatgatgctt gcttggtgtt gggg 24 23 24 DNA Homo sapiens 23 tgaagtacagggtgaggttg aagg 24 24 24 DNA Homo sapiens 24 catggtagct gcctccggtt cctg24 25 21 DNA Homo sapiens 25 atcagagtca ccaaggtgga c 21 26 25 DNA Homosapiens 26 agaaacacat cgtcctctgc acacc 25 27 25 DNA Homo sapiens 27caggctttcc tctcaccgca aacac 25 28 25 DNA Homo sapiens 28 cgtacttgactggcttgtac ctgcc 25 29 25 DNA Homo sapiens 29 tctacgtcac cagggtcatcagtgc 25 30 21 DNA Homo sapiens 30 gcctgttcac cgccttgctg t 21 31 24 DNAHomo sapiens 31 catcactgac ctctacaagt atcc 24 32 21 DNA Homo sapiens 32acggcgtggg attcatgcag g 21 33 21 DNA Homo sapiens 33 catcctctatacccgcaagc c 21 34 28 DNA Homo sapiens 34 aggttgaggc aggattaact gaggcctc28 35 22 DNA Homo sapiens 35 gatgtctgga acacagcttt gg 22 36 23 DNA Homosapiens 36 tcacagtcca gggctctgct cag 23 37 24 DNA Homo sapiens 37caaccttgcg gcttgagtct gtcg 24 38 23 DNA Homo sapiens 38 cagcagctctgacggcttgg gtc 23 39 22 DNA Homo sapiens 39 atcacctttg tggctgagac cg 2240 28 DNA Homo sapiens 40 caagggttaa aaagtaggat gaaagttc 28 41 21 DNAHomo sapiens 41 cacagtgtga cttacccgct g 21 42 22 DNA Homo sapiens 42catacaccac gtcgttccgc tg 22 43 24 DNA Homo sapiens 43 cttaaagaacccggccaaag actg 24 44 26 DNA Homo sapiens misc_feature (22)..(22) Anynucleotide of a,c,g or t 44 cgatggtcaa gttcctatcc angttg 26 45 24 DNAHomo sapiens 45 tgttgtttcc aatttccgtt ccgc 24 46 23 DNA Homo sapiens 46tctggcaccc tctggatacg cag 23 47 22 DNA Homo sapiens 47 cttagggatcagccagcgca gc 22 48 21 DNA Homo sapiens 48 gcggatgaac ctggactttg g 21 4921 DNA Homo sapiens 49 gggtgacatg gactctcagc g 21 50 24 DNA Homo sapiens50 tgggcaaggc gctgagagtc catg 24 51 24 DNA Homo sapiens 51 gctggagggcgggtctcagg gagc 24 52 31 DNA Homo sapiens 52 aggacagagg tggagccaagacgacatagc c 31 53 23 DNA Homo sapiens 53 tgaagctcag ccggaggtag aag 2354 23 DNA Homo sapiens 54 gacagactca agccgcaagg ttg 23 55 23 DNA Homosapiens 55 cagcctcagt caccgccatc cgc 23 56 23 DNA Homo sapiens 56gatacttgta gaggtcagtg atg 23 57 24 DNA Homo sapiens 57 tgggcaaggcgctgagagtc catg 24 58 24 DNA Homo sapiens 58 gctggagggc gggtctcagg gagc24 59 31 DNA Homo sapiens 59 aggacagagg tggagccaag acgacatagc c 31 60 23DNA Homo sapiens 60 cagcctcagt caccgccatc cgc 23 61 23 DNA Homo sapiens61 gatacttgta gaggtcagtg atg 23 62 21 DNA Homo sapiens 62 gtgaaggactgtgcgctgat g 21 63 21 DNA Homo sapiens 63 cgggaagtga gggcttacaa g 21 6422 DNA Homo sapiens 64 tgaggactgt gtgctgacct tg 22 65 19 DNA Homosapiens 65 tcctgtgtct ggcctaggg 19 66 22 DNA Homo sapiens 66 caggaggagtaaggggaatt gg 22 67 21 DNA Homo sapiens 67 tgcaggctac tggaatccac g 21 6820 DNA Homo sapiens 68 gggagtggcc tcatcaatgg 20 69 21 DNA Homo sapiens69 ccttgacctc actgctgctg a 21 70 21 DNA Homo sapiens 70 tcggtggcagcctcaagatt g 21 71 22 DNA Homo sapiens 71 agacatcaat gggcaggcat gg 22 7220 DNA Homo sapiens 72 ttggggagct ggagagcaac 20 73 20 DNA Homo sapiens73 atccaggact ccacccagca 20 74 34 DNA Homo sapiens 74 gcggccgctagtccacattg cagggctcct gact 34 75 23 DNA Homo sapiens 75 gctagcctgtgctcaccatg agg 23 76 46 DNA Homo sapiens 76 gctaccatgg cccaaggtagggaaagcccc tgtggccact ggagtt 46 77 44 DNA Homo sapiens CDS (30)..(44) 77ttttgtctaa ccctggctgc cccattgca ggg cta gaa gtg gcc 44 Gly Leu Glu ValAla 1 5 78 5 PRT Homo sapiens 78 Gly Leu Glu Val Ala 1 5 79 46 DNA Homosapiens CDS (1)..(15) 79 ttc gtg gtt gaa act ggtaagaacc ccagctggctcacaggggct g 46 Phe Val Val Glu Thr 1 5 80 5 PRT Homo sapiens 80 Phe ValVal Glu Thr 1 5 81 44 DNA Homo sapiens CDS (30)..(44) 81 tggagggcctcagctctact tccctccta gga ccg ctg cca gac 44 Gly Pro Leu Pro Asp 1 5 82 5PRT Homo sapiens 82 Gly Pro Leu Pro Asp 1 5 83 46 DNA Homo sapiens CDS(1)..(15) 83 aat ccc tgg tgc cca ggtaaggctg ggtgcccagg cggtgcctcc t 46Asn Pro Trp Cys Pro 1 5 84 5 PRT Homo sapiens 84 Asn Pro Trp Cys Pro 1 585 44 DNA Homo sapiens CDS (30)..(44) 85 tgcttcgtgc cctcccactc tggttcctagag gat gct gtc tac 44 Glu Asp Ala Val Tyr 1 5 86 5 PRT Homo sapiens 86Glu Asp Ala Val Tyr 1 5 87 45 DNA Homo sapiens CDS (1)..(15) 87 tgg aactat gga cag gtgagtctca gccctgctta tggcccatcc 45 Trp Asn Tyr Gly Gln 1 588 5 PRT Homo sapiens 88 Trp Asn Tyr Gly Gln 1 5 89 45 DNA Homo sapiensCDS (31)..(45) 89 tgccttccct ctctgcctct ccccccgaag ttt gaa gac aaa atc45 Phe Glu Asp Lys Ile 1 5 90 5 PRT Homo sapiens 90 Phe Glu Asp Lys Ile1 5 91 45 DNA Homo sapiens CDS (1)..(15) 91 gtg gtg tgt gcc atggtgaggtccc tggcgtgccc ggggaggagg 45 Val Val Cys Ala Met 1 5 92 5 PRTHomo sapiens 92 Val Val Cys Ala Met 1 5 93 45 DNA Homo sapiens CDS(31)..(45) 93 ctcacacttc tctatatggc ttctcttcag atc aac agc aat gat 45Ile Asn Ser Asn Asp 1 5 94 5 PRT Homo sapiens 94 Ile Asn Ser Asn Asp 1 595 46 DNA Homo sapiens CDS (1)..(15) 95 gcc gtc atg tgc aca ggtaggaggtagaaaggacc tcacaaaaag g 46 Ala Val Met Cys Thr 1 5 96 5 PRT Homo sapiens96 Ala Val Met Cys Thr 1 5 97 44 DNA Homo sapiens CDS (30)..(44) 97acaggtgatt tttttgtgcc ctttttgca gtg atg agg tgt ctg 44 Val Met Arg CysLeu 1 5 98 5 PRT Homo sapiens 98 Val Met Arg Cys Leu 1 5 99 44 DNA Homosapiens CDS (1)..(15) 99 aag gat act atc tgg tgagaaacaa cctctcaacctatttctag 44 Lys Asp Thr Ile Trp 1 5 100 5 PRT Homo sapiens 100 Lys AspThr Ile Trp 1 5 101 46 DNA Homo sapiens CDS (32)..(46) 101 caacgtctcccttggctctg tttgatacag g aac ttc cat gtc tgg 46 Asn Phe His Val Trp 1 5102 5 PRT Homo sapiens 102 Asn Phe His Val Trp 1 5 103 46 DNA Homosapiens CDS (1)..(15) 103 cag gag atg agc aac ggtgaggctc tccagaagaaaggcaggccc c 46 Gln Glu Met Ser Asn 1 5 104 5 PRT Homo sapiens 104 GlnGlu Met Ser Asn 1 5 105 44 DNA Homo sapiens CDS (30)..(44) 105gcccaccgag gctcccctgt tctccttca ggc gtc tac tgc tgt 44 Gly Val Tyr CysCys 1 5 106 5 PRT Homo sapiens 106 Gly Val Tyr Cys Cys 1 5 107 46 DNAHomo sapiens CDS (1)..(15) 107 tac aag tat gaa gaa ggttagtaag caagccagccctactcagag c 46 Tyr Lys Tyr Glu Glu 1 5 108 5 PRT Homo sapiens 108 TyrLys Tyr Glu Glu 1 5 109 44 DNA Homo sapiens CDS (30)..(44) 109cagctggtgc tgtgctctcc ccaacttca gga tcc ctc cag gag 44 Gly Ser Leu GlnGlu 1 5 110 5 PRT Homo sapiens 110 Gly Ser Leu Gln Glu 1 5 111 46 DNAHomo sapiens CDS (1)..(15) 111 ctc tct cct aaa gaa ggtacgcatg tgcacagtttgtgtacgcag a 46 Leu Ser Pro Lys Glu 1 5 112 5 PRT Homo sapiens 112 LeuSer Pro Lys Glu 1 5 113 44 DNA Homo sapiens CDS (30)..(44) 113tctcacccca tccttgtgtt ctttcttta gca aag acc tac ccc 44 Ala Lys Thr TyrPro 1 5 114 5 PRT Homo sapiens 114 Ala Lys Thr Tyr Pro 1 5 115 45 DNAHomo sapiens CDS (1)..(15) 115 agc atc acg att aat gtaggcagga gtcctgcaaatggcttgtgg 45 Ser Ile Thr Ile Asn 1 5 116 5 PRT Homo sapiens 116 Ser IleThr Ile Asn 1 5 117 45 DNA Homo sapiens CDS (31)..(45) 117 taattctccttcccctcctg gtctgtttag gtt cta gga gca gcc 45 Val Leu Gly Ala Ala 1 5 1185 PRT Homo sapiens 118 Val Leu Gly Ala Ala 1 5 119 44 DNA Homo sapiensCDS (1)..(15) 119 cag cag aaa gtc ttg taagtgctgc aagtgctcag ccttctcct 44Gln Gln Lys Val Leu 1 5 120 5 PRT Homo sapiens 120 Gln Gln Lys Val Leu 15 121 46 DNA Homo sapiens CDS (32)..(46) 121 ttttctgaca tgctccattctctgttgcag c ctt gga gtc ctc aaa 46 Leu Gly Val Leu Lys 1 5 122 5 PRTHomo sapiens 122 Leu Gly Val Leu Lys 1 5 123 32 DNA Homo sapiens 123cggtggacta atggctcgtc tagtgttgac ca 32 124 529 DNA Homo sapiens 124agggttctgt ttgcatttaa tgttcaggaa gtgacttttc aggataaaag agaggccttg 60gttacctagc agggttctcg gttcaatgag gtgctttaag ctgtgagcag agtcttggga 120ccctgatgct ccttccagca gccccctgca tccctctggc gggcccatca gctcgcttct 180ccctcctgac ttctccacca cagcacagca cctccctggg aatgccctat tgctcaagag 240ctatcaaagg cccacacggc ataaggctgt gacagttcat cagcctccac acctcctttc 300aattcagcaa cactgccaag aaaaacctga gggcaagtga gcaaaccagt tgtgatctgc 360tcggtaatca ggtggcagtg cagcagtcca gccccgcttc ggttctcctg gaggcttcca 420atggaagggg aagtagacac tctggcacca gtttgctgaa gctccagacc gcccagctgt 480tctgtgggga gcatcccagg aaccggaggc agctaccatg gcccaaggt 529 125 793 DNAHomo sapiens CDS (1)..(15) 125 gta gac ttt gca tta taaattctgg aacaacgcgccagacgtgtg aatttcaagc 55 Val Asp Phe Ala Leu 1 5 ttcaggaaaa ggagcaagttcaaatgcaag ctgcgccatt ccccaccaca acagaggctt 115 cacagggctc cagcaagagccacagagggg atgacgtgtt cattttctgt ctctcctgac 175 tccactagaa atttaagctccatgagggca aagactttgc tttgtttact acccctatac 235 tcagaaccat ttctggcatatgctaggcac tcaacaaata ttttttgaat gaatgaatag 295 gagactccag catccagagaacaggtagga aatgtctatg gatggatatt tccctggacc 355 atttgcacag ctcccctggactcttttcag ggcccaggga ttccactgtg tcccatccag 415 agattccagg attccagtcacctatccaga agcgtgattt ggcacagagg tcagaggata 475 ctggtaggac ttggccatgacttaactgcc ccctgcccca gatatccagg aaagaaaaag 535 acaggctgaa cagctcactgttgttttgtt gttgcgaaag ctaattccct agtatgaata 595 aacttcagac cttgctctcctttgcctcat gtagtcatca ctttcctatc tgttccttgg 655 atagctgcag tctccattcattcaaaaaag tcatttattg agtgcctagc atatgccaga 715 agtggttctg agtgtaggggtacaaagtaa acaaagcaaa gtccctgccc tcatggagcg 775 cacatttctc agtggagg 793126 5 PRT Homo sapiens 126 Val Asp Phe Ala Leu 1 5 127 2317 DNA Homosapiens 127 accacatggt gggaatggca accttgcggc ttgagtctgt cgacctgcagagctccagga 60 acaacaagga gcaccacacg caggagatgg gcgtcaagcg gctcactgtgcgccgcggcc 120 agcccttcta cctccggctg agcttcagcc gacccttcca gtcccagaacgaccacatca 180 cctttgtggc tgagaccgga cccaagccgt cagagctgct ggggacccgagccacattct 240 tcctcacccg ggtccagccc gggaatgtct ggagcgcttc tgatttcaccattgactcca 300 actctctcca agtttccctt ttcacaccag ccaatgcagt tattggccattacactctga 360 aaatagagat ctctcagggc caaggtcaca gtgtgactta cccgctgggaactttcatcc 420 tactttttaa cccttggagt ccagaggacg acgtctacct gccaagtgaaatactgctgc 480 aggagtatat catgcgagat tatggctttg tttacaaggg tcatgaaagattcatcacct 540 cctggccctg gaactacggg cagtttgaag aggacatcat agacatctgctttgagatcc 600 tgaacaagag cctgtatcac ttaaagaacc cggccaaaga ctgttcccagcggaacgacg 660 tggtgtatgt gtgcagggtg gtgagtgcca tgatcaacag caacgatgacaatggcgtgc 720 tgcaggggaa ctggggcgag gactactcca aaggggtcag tcctctggagtggaagggca 780 gtgtggccat cctacagcag tggtcagcca ggggcgggca gcctgtgaagtacggacagt 840 gctgggtctt cgcctctgtt atgtgcaccg taatgagatg cttaggtgttccaacccgtg 900 ttgtttccaa tttccgttcc gcgcacaacg tggataggaa cttgaccatcgatacgtact 960 atgaccgaaa tgccgagatg ctgtcaactc agaaacgaga caaaatatggaacttccacg 1020 tctggaatga gtgctggatg atccggaaag atctcccacc aggatacaacgggtggcagg 1080 ttctggaccc cactccccag cagaccagca gtgggctgtt ctgctgtggccctgcctctg 1140 tgaaggccat cagggaaggg gatgtccacc tggcctatga caccccttttgtgtatgccg 1200 aggtgaacgc cgatgaagtc atttggctcc ttggggatgg ccaggcccaggaaatcctgg 1260 cccacaacac cagttccatc gggaaggaga tcagcactaa gatggtggggtcagaccagc 1320 gccagagcat caccagctcc tacaagtacc cagaaggatc ccctgaggagagagctgtct 1380 tcatgaaggc ttctcggaaa atgctgggcc cccaaagagc ttctttgcccttcctggatc 1440 tcctggagtc tgggggtctt agggatcagc cagcgcagct gcagcttcacctggccagga 1500 tacccgagtg gggccaggac ctgcagctgc tgctgcgtat ccagagggtgccagacagca 1560 cccaccctcg ggggcccatc ggactggtgg tgcgcttctg tgcacaggccctgctgcatg 1620 ggggtggtac ccagaagccc ttctggaggc acacagtgcg gatgaacctggactttggga 1680 aggagacaca gtggccgctc ctcctgccct acagcaatta cagaaacaagctaacggacg 1740 aaaagctcat ccgcgtgtct ggcatcgcgg aggttgaaga gacagggaggtccatgctgg 1800 tcctaaaaga tatctgtctg gagcctcccc acttgtctat tgaggtgtctgagagggctg 1860 aggtgggcaa ggcgctgaga gtccatgtca ccctcaccaa caccttaatggtggctctga 1920 gcagctgcac gatggtgctg gaaggaagcg gcctcatcaa tgggcagatagcaaaggacc 1980 ttgggactct ggtggccgga cacaccctcc aaattcaact ggacctctacccgaccaaag 2040 ctggaccccg ccagctccag gttctcatca gcagcaacga ggtcaaggagatcaaaggct 2100 acaaggacat attcgtcact gtggctgggg ctccctgaga cccgccctccagctgccctc 2160 cctggcaccc ctgccccacc tggctccttt ctactcctgg ctatgtcgtcttggctccac 2220 ctctgtcctc tctctagcct gcctgggaat gaatgaagct ctgttagaaacaccgtgtgc 2280 tttgggaaga gacaataaag atgtctttat ttatcac 2317 128 710PRT Homo sapiens 128 Met Val Gly Met Ala Thr Leu Arg Leu Glu Ser Val AspLeu Gln Ser 1 5 10 15 Ser Arg Asn Asn Lys Glu His His Thr Gln Glu MetGly Val Lys Arg 20 25 30 Leu Thr Val Arg Arg Gly Gln Pro Phe Tyr Leu ArgLeu Ser Phe Ser 35 40 45 Arg Pro Phe Gln Ser Gln Asn Asp His Ile Thr PheVal Ala Glu Thr 50 55 60 Gly Pro Lys Pro Ser Glu Leu Leu Gly Thr Arg AlaThr Phe Phe Leu 65 70 75 80 Thr Arg Val Gln Pro Gly Asn Val Trp Ser AlaSer Asp Phe Thr Ile 85 90 95 Asp Ser Asn Ser Leu Gln Val Ser Leu Phe ThrPro Ala Asn Ala Val 100 105 110 Ile Gly His Tyr Thr Leu Lys Ile Glu IleSer Gln Gly Gln Gly His 115 120 125 Ser Val Thr Tyr Pro Leu Gly Thr PheIle Leu Leu Phe Asn Pro Trp 130 135 140 Ser Pro Glu Asp Asp Val Tyr LeuPro Ser Glu Ile Leu Leu Gln Glu 145 150 155 160 Tyr Ile Met Arg Asp TyrGly Phe Val Tyr Lys Gly His Glu Arg Phe 165 170 175 Ile Thr Ser Trp ProTrp Asn Tyr Gly Gln Phe Glu Glu Asp Ile Ile 180 185 190 Asp Ile Cys PheGlu Ile Leu Asn Lys Ser Leu Tyr His Leu Lys Asn 195 200 205 Pro Ala LysAsp Cys Ser Gln Arg Asn Asp Val Val Tyr Val Cys Arg 210 215 220 Val ValSer Ala Met Ile Asn Ser Asn Asp Asp Asn Gly Val Leu Gln 225 230 235 240Gly Asn Trp Gly Glu Asp Tyr Ser Lys Gly Val Ser Pro Leu Glu Trp 245 250255 Lys Gly Ser Val Ala Ile Leu Gln Gln Trp Ser Ala Arg Gly Gly Gln 260265 270 Pro Val Lys Tyr Gly Gln Cys Trp Val Phe Ala Ser Val Met Cys Thr275 280 285 Val Met Arg Cys Leu Gly Val Pro Thr Arg Val Val Ser Asn PheArg 290 295 300 Ser Ala His Asn Val Asp Arg Asn Leu Thr Ile Asp Thr TyrTyr Asp 305 310 315 320 Arg Asn Ala Glu Met Leu Ser Thr Gln Lys Arg AspLys Ile Trp Asn 325 330 335 Phe His Val Trp Asn Glu Cys Trp Met Ile ArgLys Asp Leu Pro Pro 340 345 350 Gly Tyr Asn Gly Trp Gln Val Leu Asp ProThr Pro Gln Gln Thr Ser 355 360 365 Ser Gly Leu Phe Cys Cys Gly Pro AlaSer Val Lys Ala Ile Arg Glu 370 375 380 Gly Asp Val His Leu Ala Tyr AspThr Pro Phe Val Tyr Ala Glu Val 385 390 395 400 Asn Ala Asp Glu Val IleTrp Leu Leu Gly Asp Gly Gln Ala Gln Glu 405 410 415 Ile Leu Ala His AsnThr Ser Ser Ile Gly Lys Glu Ile Ser Thr Lys 420 425 430 Met Val Gly SerAsp Gln Arg Gln Ser Ile Thr Ser Ser Tyr Lys Tyr 435 440 445 Pro Glu GlySer Pro Glu Glu Arg Ala Val Phe Met Lys Ala Ser Arg 450 455 460 Lys MetLeu Gly Pro Gln Arg Ala Ser Leu Pro Phe Leu Asp Leu Leu 465 470 475 480Glu Ser Gly Gly Leu Arg Asp Gln Pro Ala Gln Leu Gln Leu His Leu 485 490495 Ala Arg Ile Pro Glu Trp Gly Gln Asp Leu Gln Leu Leu Leu Arg Ile 500505 510 Gln Arg Val Pro Asp Ser Thr His Pro Arg Gly Pro Ile Gly Leu Val515 520 525 Val Arg Phe Cys Ala Gln Ala Leu Leu His Gly Gly Gly Thr GlnLys 530 535 540 Pro Phe Trp Arg His Thr Val Arg Met Asn Leu Asp Phe GlyLys Glu 545 550 555 560 Thr Gln Trp Pro Leu Leu Leu Pro Tyr Ser Asn TyrArg Asn Lys Leu 565 570 575 Thr Asp Glu Lys Leu Ile Arg Val Ser Gly IleAla Glu Val Glu Glu 580 585 590 Thr Gly Arg Ser Met Leu Val Leu Lys AspIle Cys Leu Glu Pro Pro 595 600 605 His Leu Ser Ile Glu Val Ser Glu ArgAla Glu Val Gly Lys Ala Leu 610 615 620 Arg Val His Val Thr Leu Thr AsnThr Leu Met Val Ala Leu Ser Ser 625 630 635 640 Cys Thr Met Val Leu GluGly Ser Gly Leu Ile Asn Gly Gln Ile Ala 645 650 655 Lys Asp Leu Gly ThrLeu Val Ala Gly His Thr Leu Gln Ile Gln Leu 660 665 670 Asp Leu Tyr ProThr Lys Ala Gly Pro Arg Gln Leu Gln Val Leu Ile 675 680 685 Ser Ser AsnGlu Val Lys Glu Ile Lys Gly Tyr Lys Asp Ile Phe Val 690 695 700 Thr ValAla Gly Ala Pro 705 710 129 2312 DNA Homo sapiens 129 gggagatggatcaggtggca accttgcggc ttgagtctgt cgacctgcag agctccagga 60 acaacaaggagcaccacacg caggagatgg gcgtcaagcg gctcactgtg cgccgcggcc 120 agcccttctacctccggctg agcttcagcc gacccttcca gtcccagaac gaccacatca 180 cctttgtggctgagaccgga cccaagccgt cagagctgct ggggacccga gccacattct 240 tcctcacccgggtccagccc gggaatgtct ggagcgcttc tgatttcacc attgactcca 300 actctctccaagtttccctt ttcacaccag ccaatgcagt tattggccat tacactctga 360 aaatagagatctctcagggc caaggtcaca gtgtgactta cccgctggga actttcatcc 420 tactttttaacccttggagt ccagaggacg acgtctacct gccaagtgaa atactgctgc 480 aggagtatatcatgcgagat tatggctttg tttacaaggg tcatgaaaga ttcatcacct 540 cctggccctggaactacggg cagtttgaag aggacatcat agacatctgc tttgagatcc 600 tgaacaagagcctgtatcac ttaaagaacc cggccaaaga ctgttcccag cggaacgacg 660 tggtgtatgtgtgcagggtg gtgagtgcca tgatcaacag caacgatgac aatggcgtgc 720 tgcaggggaactggggcgag gactactcca aaggggtcag tcctctggag tggaagggca 780 gtgtggccatcctacagcag tggtcagcca ggggcgggca gcctgtgaag tacggacagt 840 gctgggtcttcgcctctgtt atgtgcaccg taatgagatg cttaggtgtt ccaacccgtg 900 ttgtttccaatttccgttcc gcgcacaacg tggataggaa cttgaccatc gatacgtact 960 atgaccgaaatgccgagatg ctgtcaactc agaaacgaga caaaatatgg aacttccacg 1020 tctggaatgagtgctggatg atccggaaag atctcccacc aggatacaac gggtggcagg 1080 ttctggaccccactccccag cagaccagca gtgggctgtt ctgctgtggc cctgcctctg 1140 tgaaggccatcagggaaggg gatgtccacc tggcctatga cacccctttt gtgtatgccg 1200 aggtgaacgccgatgaagtc atttggctcc ttggggatgg ccaggcccag gaaatcctgg 1260 cccacaacaccagttccatc gggaaggaga tcagcactaa gatggtgggg tcagaccagc 1320 gccagagcatcaccagctcc tacaagtacc cagaaggatc ccctgaggag agagctgtct 1380 tcatgaaggcttctcggaaa atgctgggcc cccaaagagc ttctttgccc ttcctggatc 1440 tcctggagtctgggggtctt agggatcagc cagcgcagct gcagcttcac ctggccagga 1500 tacccgagtggggccaggac ctgcagctgc tgctgcgtat ccagagggtg ccagacagca 1560 cccaccctcgggggcccatc ggactggtgg tgcgcttctg tgcacaggcc ctgctgcatg 1620 ggggtggtacccagaagccc ttctggaggc acacagtgcg gatgaacctg gactttggga 1680 aggagacacagtggccgctc ctcctgccct acagcaatta cagaaacaag ctaacggacg 1740 aaaagctcatccgcgtgtct ggcatcgcgg aggttgaaga gacagggagg tccatgctgg 1800 tcctaaaagatatctgtctg gagcctcccc acttgtctat tgaggtgtct gagagggctg 1860 aggtgggcaaggcgctgaga gtccatgtca ccctcaccaa caccttaatg gtggctctga 1920 gcagctgcacgatggtgctg gaaggaagcg gcctcatcaa tgggcagata gcaaaggacc 1980 ttgggactctggtggccgga cacaccctcc aaattcaact ggacctctac ccgaccaaag 2040 ctggaccccgccagctccag gttctcatca gcagcaacga ggtcaaggag atcaaaggct 2100 acaaggacatattcgtcact gtggctgggg ctccctgaga cccgccctcc agctgccctc 2160 cctggcacccctgccccacc tggctccttt ctactcctgg ctatgtcgtc ttggctccac 2220 ctctgtcctctctctagcct gcctgggaat gaatgaagct ctgttagaaa caccgtgtgc 2280 tttgggaagagacaataaag atgtctttat tt 2312 130 710 PRT Homo sapiens 130 Met Asp GlnVal Ala Thr Leu Arg Leu Glu Ser Val Asp Leu Gln Ser 1 5 10 15 Ser ArgAsn Asn Lys Glu His His Thr Gln Glu Met Gly Val Lys Arg 20 25 30 Leu ThrVal Arg Arg Gly Gln Pro Phe Tyr Leu Arg Leu Ser Phe Ser 35 40 45 Arg ProPhe Gln Ser Gln Asn Asp His Ile Thr Phe Val Ala Glu Thr 50 55 60 Gly ProLys Pro Ser Glu Leu Leu Gly Thr Arg Ala Thr Phe Phe Leu 65 70 75 80 ThrArg Val Gln Pro Gly Asn Val Trp Ser Ala Ser Asp Phe Thr Ile 85 90 95 AspSer Asn Ser Leu Gln Val Ser Leu Phe Thr Pro Ala Asn Ala Val 100 105 110Ile Gly His Tyr Thr Leu Lys Ile Glu Ile Ser Gln Gly Gln Gly His 115 120125 Ser Val Thr Tyr Pro Leu Gly Thr Phe Ile Leu Leu Phe Asn Pro Trp 130135 140 Ser Pro Glu Asp Asp Val Tyr Leu Pro Ser Glu Ile Leu Leu Gln Glu145 150 155 160 Tyr Ile Met Arg Asp Tyr Gly Phe Val Tyr Lys Gly His GluArg Phe 165 170 175 Ile Thr Ser Trp Pro Trp Asn Tyr Gly Gln Phe Glu GluAsp Ile Ile 180 185 190 Asp Ile Cys Phe Glu Ile Leu Asn Lys Ser Leu TyrHis Leu Lys Asn 195 200 205 Pro Ala Lys Asp Cys Ser Gln Arg Asn Asp ValVal Tyr Val Cys Arg 210 215 220 Val Val Ser Ala Met Ile Asn Ser Asn AspAsp Asn Gly Val Leu Gln 225 230 235 240 Gly Asn Trp Gly Glu Asp Tyr SerLys Gly Val Ser Pro Leu Glu Trp 245 250 255 Lys Gly Ser Val Ala Ile LeuGln Gln Trp Ser Ala Arg Gly Gly Gln 260 265 270 Pro Val Lys Tyr Gly GlnCys Trp Val Phe Ala Ser Val Met Cys Thr 275 280 285 Val Met Arg Cys LeuGly Val Pro Thr Arg Val Val Ser Asn Phe Arg 290 295 300 Ser Ala His AsnVal Asp Arg Asn Leu Thr Ile Asp Thr Tyr Tyr Asp 305 310 315 320 Arg AsnAla Glu Met Leu Ser Thr Gln Lys Arg Asp Lys Ile Trp Asn 325 330 335 PheHis Val Trp Asn Glu Cys Trp Met Ile Arg Lys Asp Leu Pro Pro 340 345 350Gly Tyr Asn Gly Trp Gln Val Leu Asp Pro Thr Pro Gln Gln Thr Ser 355 360365 Ser Gly Leu Phe Cys Cys Gly Pro Ala Ser Val Lys Ala Ile Arg Glu 370375 380 Gly Asp Val His Leu Ala Tyr Asp Thr Pro Phe Val Tyr Ala Glu Val385 390 395 400 Asn Ala Asp Glu Val Ile Trp Leu Leu Gly Asp Gly Gln AlaGln Glu 405 410 415 Ile Leu Ala His Asn Thr Ser Ser Ile Gly Lys Glu IleSer Thr Lys 420 425 430 Met Val Gly Ser Asp Gln Arg Gln Ser Ile Thr SerSer Tyr Lys Tyr 435 440 445 Pro Glu Gly Ser Pro Glu Glu Arg Ala Val PheMet Lys Ala Ser Arg 450 455 460 Lys Met Leu Gly Pro Gln Arg Ala Ser LeuPro Phe Leu Asp Leu Leu 465 470 475 480 Glu Ser Gly Gly Leu Arg Asp GlnPro Ala Gln Leu Gln Leu His Leu 485 490 495 Ala Arg Ile Pro Glu Trp GlyGln Asp Leu Gln Leu Leu Leu Arg Ile 500 505 510 Gln Arg Val Pro Asp SerThr His Pro Arg Gly Pro Ile Gly Leu Val 515 520 525 Val Arg Phe Cys AlaGln Ala Leu Leu His Gly Gly Gly Thr Gln Lys 530 535 540 Pro Phe Trp ArgHis Thr Val Arg Met Asn Leu Asp Phe Gly Lys Glu 545 550 555 560 Thr GlnTrp Pro Leu Leu Leu Pro Tyr Ser Asn Tyr Arg Asn Lys Leu 565 570 575 ThrAsp Glu Lys Leu Ile Arg Val Ser Gly Ile Ala Glu Val Glu Glu 580 585 590Thr Gly Arg Ser Met Leu Val Leu Lys Asp Ile Cys Leu Glu Pro Pro 595 600605 His Leu Ser Ile Glu Val Ser Glu Arg Ala Glu Val Gly Lys Ala Leu 610615 620 Arg Val His Val Thr Leu Thr Asn Thr Leu Met Val Ala Leu Ser Ser625 630 635 640 Cys Thr Met Val Leu Glu Gly Ser Gly Leu Ile Asn Gly GlnIle Ala 645 650 655 Lys Asp Leu Gly Thr Leu Val Ala Gly His Thr Leu GlnIle Gln Leu 660 665 670 Asp Leu Tyr Pro Thr Lys Ala Gly Pro Arg Gln LeuGln Val Leu Ile 675 680 685 Ser Ser Asn Glu Val Lys Glu Ile Lys Gly TyrLys Asp Ile Phe Val 690 695 700 Thr Val Ala Gly Ala Pro 705 710 131 719PRT Homo sapiens 131 Ala Gln Gly Leu Glu Val Ala Leu Thr Asp Leu Gln SerSer Arg Asn 1 5 10 15 Asn Val Arg His His Thr Glu Glu Ile Thr Val AspHis Leu Leu Val 20 25 30 Arg Arg Gly Gln Ala Phe Asn Leu Thr Leu Tyr PheArg Asn Arg Ser 35 40 45 Phe Gln Pro Gly Leu Asp Asn Ile Ile Phe Val ValGlu Thr Gly Pro 50 55 60 Leu Pro Asp Leu Ala Leu Gly Thr Arg Ala Val PheSer Leu Ala Arg 65 70 75 80 His His Ser Pro Ser Pro Trp Ile Ala Trp LeuGlu Thr Asn Gly Ala 85 90 95 Thr Ser Thr Glu Val Ser Leu Cys Ala Pro ProThr Ala Ala Val Gly 100 105 110 Arg Tyr Leu Leu Lys Ile His Ile Asp SerPhe Gln Gly Ser Val Thr 115 120 125 Ala Tyr Gln Leu Gly Glu Phe Ile LeuLeu Phe Asn Pro Trp Cys Pro 130 135 140 Glu Asp Ala Val Tyr Leu Asp SerGlu Pro Gln Arg Gln Glu Tyr Val 145 150 155 160 Met Asn Asp Tyr Gly PheIle Tyr Gln Gly Ser Lys Asn Trp Ile Arg 165 170 175 Pro Cys Pro Trp AsnTyr Gly Gln Phe Glu Asp Lys Ile Ile Asp Ile 180 185 190 Cys Leu Lys LeuLeu Asp Lys Ser Leu His Phe Gln Thr Asp Pro Ala 195 200 205 Thr Asp CysAla Leu Arg Gly Ser Pro Val Tyr Val Ser Arg Val Val 210 215 220 Cys AlaMet Ile Asn Ser Asn Asp Asp Asn Gly Val Leu Asn Gly Asn 225 230 235 240Trp Ser Glu Asn Tyr Thr Asp Gly Ala Asn Pro Ala Glu Trp Thr Gly 245 250255 Ser Val Ala Ile Leu Lys Gln Trp Asn Ala Thr Gly Cys Gln Pro Val 260265 270 Arg Tyr Gly Gln Cys Trp Val Phe Ala Ala Val Met Cys Thr Val Met275 280 285 Arg Cys Leu Gly Ile Pro Thr Arg Val Ile Thr Asn Phe Asp SerGly 290 295 300 His Asp Thr Asp Gly Asn Leu Ile Ile Asp Glu Tyr Tyr AspAsn Thr 305 310 315 320 Gly Arg Ile Leu Gly Asn Lys Lys Lys Asp Thr IleTrp Asn Phe His 325 330 335 Val Trp Asn Glu Cys Trp Met Ala Arg Lys AspLeu Pro Pro Gly Tyr 340 345 350 Gly Gly Trp Gln Val Leu Asp Ala Thr ProGln Glu Met Ser Asn Gly 355 360 365 Val Tyr Cys Cys Gly Pro Ala Ser ValArg Ala Ile Lys Glu Gly Glu 370 375 380 Val Asp Leu Asn Tyr Asp Thr ProPhe Val Phe Ser Met Val Asn Ala 385 390 395 400 Asp Cys Met Ser Trp LeuVal Gln Gly Gly Lys Glu Gln Lys Leu His 405 410 415 Gln Asp Thr Ser SerVal Gly Asn Phe Ile Ser Thr Lys Ser Ile Gln 420 425 430 Ser Asp Glu ArgAsp Asp Ile Thr Glu Asn Tyr Lys Tyr Glu Glu Gly 435 440 445 Ser Leu GlnGlu Arg Gln Val Phe Leu Lys Ala Leu Gln Lys Leu Lys 450 455 460 Ala ArgSer Phe His Gly Ser Gln Arg Gly Ala Glu Leu Gln Pro Ser 465 470 475 480Arg Pro Thr Ser Leu Ser Gln Asp Ser Pro Arg Ser Leu His Thr Pro 485 490495 Ser Leu Arg Pro Ser Asp Val Val Gln Val Ser Leu Lys Phe Lys Leu 500505 510 Leu Asp Pro Pro Asn Met Gly Gln Asp Ile Cys Phe Val Leu Leu Ala515 520 525 Leu Asn Met Ser Ser Gln Phe Lys Asp Leu Lys Val Asn Leu SerAla 530 535 540 Gln Ser Leu Leu His Asp Gly Ser Pro Leu Ser Pro Phe TrpGln Asp 545 550 555 560 Thr Ala Phe Ile Thr Leu Ser Pro Lys Glu Ala LysThr Tyr Pro Cys 565 570 575 Lys Ile Ser Tyr Ser Gln Tyr Ser Gln Tyr LeuSer Thr Asp Lys Leu 580 585 590 Ile Arg Ile Ser Ala Leu Gly Glu Glu LysSer Ser Pro Glu Lys Ile 595 600 605 Leu Val Asn Lys Ile Ile Thr Leu SerTyr Pro Ser Ile Thr Ile Asn 610 615 620 Val Leu Gly Ala Ala Val Val AsnGln Pro Leu Ser Ile Gln Val Ile 625 630 635 640 Phe Ser Asn Pro Leu SerGlu Gln Val Glu Asp Cys Val Leu Thr Val 645 650 655 Glu Gly Ser Gly LeuPhe Lys Lys Gln Gln Lys Val Phe Leu Gly Val 660 665 670 Leu Lys Pro GlnHis Gln Ala Ser Ile Ile Leu Glu Thr Val Pro Phe 675 680 685 Lys Ser GlyGln Arg Gln Ile Gln Ala Asn Met Arg Ser Asn Lys Phe 690 695 700 Lys AspIle Lys Gly Tyr Arg Asn Val Tyr Val Asp Phe Ala Leu 705 710 715 132 709PRT Homo sapiens 132 Val Gly Met Ala Thr Leu Arg Leu Glu Ser Val Asp LeuGln Ser Ser 1 5 10 15 Arg Asn Asn Lys Glu His His Thr Gln Glu Met GlyVal Lys Arg Leu 20 25 30 Thr Val Arg Arg Gly Gln Pro Phe Tyr Leu Arg LeuSer Phe Ser Arg 35 40 45 Pro Phe Gln Ser Gln Asn Asp His Ile Thr Phe ValAla Glu Thr Gly 50 55 60 Pro Lys Pro Ser Glu Leu Leu Gly Thr Arg Ala ThrPhe Phe Leu Thr 65 70 75 80 Arg Val Gln Pro Gly Asn Val Trp Ser Ala SerAsp Phe Thr Ile Asp 85 90 95 Ser Asn Ser Leu Gln Val Ser Leu Phe Thr ProAla Asn Ala Val Ile 100 105 110 Gly His Tyr Thr Leu Lys Ile Glu Ile SerGln Gly Gln Gly His Ser 115 120 125 Val Thr Tyr Pro Leu Gly Thr Phe IleLeu Leu Phe Asn Pro Trp Ser 130 135 140 Pro Glu Asp Asp Val Tyr Leu ProSer Glu Ile Leu Leu Gln Glu Tyr 145 150 155 160 Ile Met Arg Asp Tyr GlyPhe Val Tyr Lys Gly His Glu Arg Phe Ile 165 170 175 Thr Ser Trp Pro TrpAsn Tyr Gly Gln Phe Glu Glu Asp Ile Ile Asp 180 185 190 Ile Cys Phe GluIle Leu Asn Lys Ser Leu Tyr His Leu Lys Asn Pro 195 200 205 Ala Lys AspCys Ser Gln Arg Asn Asp Val Val Tyr Val Cys Arg Val 210 215 220 Val SerAla Met Ile Asn Ser Asn Asp Asp Asn Gly Val Leu Gln Gly 225 230 235 240Asn Trp Gly Glu Asp Tyr Ser Lys Gly Val Ser Pro Leu Glu Trp Lys 245 250255 Gly Ser Val Ala Ile Leu Gln Gln Trp Ser Ala Arg Gly Gly Gln Pro 260265 270 Val Lys Tyr Gly Gln Cys Trp Val Phe Ala Ser Val Met Cys Thr Val275 280 285 Met Arg Cys Leu Gly Val Pro Thr Arg Val Val Ser Asn Phe ArgSer 290 295 300 Ala His Asn Val Asp Arg Asn Leu Thr Ile Asp Thr Tyr TyrAsp Arg 305 310 315 320 Asn Ala Glu Met Leu Ser Thr Gln Lys Arg Asp LysIle Trp Asn Phe 325 330 335 His Val Trp Asn Glu Cys Trp Met Ile Arg LysAsp Leu Pro Pro Gly 340 345 350 Tyr Asn Gly Trp Gln Val Leu Asp Pro ThrPro Gln Gln Thr Ser Ser 355 360 365 Gly Leu Phe Cys Cys Gly Pro Ala SerVal Lys Ala Ile Arg Glu Gly 370 375 380 Asp Val His Leu Ala Tyr Asp ThrPro Phe Val Tyr Ala Glu Val Asn 385 390 395 400 Ala Asp Glu Val Ile TrpLeu Leu Gly Asp Gly Gln Ala Gln Glu Ile 405 410 415 Leu Ala His Asn ThrSer Ser Ile Gly Lys Glu Ile Ser Thr Lys Met 420 425 430 Val Gly Ser AspGln Arg Gln Ser Ile Thr Ser Ser Tyr Lys Tyr Pro 435 440 445 Glu Gly SerPro Glu Glu Arg Ala Val Phe Met Lys Ala Ser Arg Lys 450 455 460 Met LeuGly Pro Gln Arg Ala Ser Leu Pro Phe Leu Asp Leu Leu Glu 465 470 475 480Ser Gly Gly Leu Arg Asp Gln Pro Ala Gln Leu Gln Leu His Leu Ala 485 490495 Arg Ile Pro Glu Trp Gly Gln Asp Leu Gln Leu Leu Leu Arg Ile Gln 500505 510 Arg Val Pro Asp Ser Thr His Pro Arg Gly Pro Ile Gly Leu Val Val515 520 525 Arg Phe Cys Ala Gln Ala Leu Leu His Gly Gly Gly Thr Gln LysPro 530 535 540 Phe Trp Arg His Thr Val Arg Met Asn Leu Asp Phe Gly LysGlu Thr 545 550 555 560 Gln Trp Pro Leu Leu Leu Pro Tyr Ser Asn Tyr ArgAsn Lys Leu Thr 565 570 575 Asp Glu Lys Leu Ile Arg Val Ser Gly Ile AlaGlu Val Glu Glu Thr 580 585 590 Gly Arg Ser Met Leu Val Leu Lys Asp IleCys Leu Glu Pro Pro His 595 600 605 Leu Ser Ile Glu Val Ser Glu Arg AlaGlu Val Gly Lys Ala Leu Arg 610 615 620 Val His Val Thr Leu Thr Asn ThrLeu Met Val Ala Leu Ser Ser Cys 625 630 635 640 Thr Met Val Leu Glu GlySer Gly Leu Ile Asn Gly Gln Ile Ala Lys 645 650 655 Asp Leu Gly Thr LeuVal Ala Gly His Thr Leu Gln Ile Gln Leu Asp 660 665 670 Leu Tyr Pro ThrLys Ala Gly Pro Arg Gln Leu Gln Val Leu Ile Ser 675 680 685 Ser Asn GluVal Lys Glu Ile Lys Gly Tyr Lys Asp Ile Phe Val Thr 690 695 700 Val AlaGly Ala Pro 705 133 690 PRT Homo sapiens 133 Gly Gln Ala Leu Gly Ile LysSer Cys Asp Phe Gln Ala Ala Arg Asn 1 5 10 15 Asn Glu Glu His His ThrLys Ala Leu Ser Ser Arg Arg Leu Phe Val 20 25 30 Arg Arg Gly Gln Pro PheThr Ile Ile Leu Tyr Phe Arg Ala Pro Val 35 40 45 Arg Ala Phe Leu Pro AlaLeu Lys Lys Val Ala Leu Thr Ala Gln Thr 50 55 60 Gly Glu Gln Pro Ser LysIle Asn Arg Thr Gln Ala Thr Phe Pro Ile 65 70 75 80 Ser Ser Leu Gly AspArg Lys Trp Trp Ser Ala Val Val Glu Glu Arg 85 90 95 Asp Ala Gln Ser TrpThr Ile Ser Val Thr Thr Pro Ala Asp Ala Val 100 105 110 Ile Gly His TyrSer Leu Leu Leu Gln Val Ser Gly Arg Lys Gln Leu 115 120 125 Leu Leu GlyGln Phe Thr Leu Leu Phe Asn Pro Trp Asn Arg Glu Asp 130 135 140 Ala ValPhe Leu Lys Asn Glu Ala Gln Arg Met Glu Tyr Leu Leu Asn 145 150 155 160Gln Asn Gly Leu Ile Tyr Leu Gly Thr Ala Asp Cys Ile Gln Ala Glu 165 170175 Ser Trp Asp Phe Gly Gln Phe Glu Gly Asp Val Ile Asp Leu Ser Leu 180185 190 Arg Leu Leu Ser Lys Asp Lys Gln Val Glu Lys Trp Ser Gln Pro Val195 200 205 His Val Ala Arg Val Leu Gly Ala Leu Leu His Phe Leu Lys GluGln 210 215 220 Arg Val Leu Pro Thr Pro Gln Thr Gln Ala Thr Gln Glu GlyAla Leu 225 230 235 240 Leu Asn Lys Arg Arg Gly Ser Val Pro Ile Leu ArgGln Trp Leu Thr 245 250 255 Gly Arg Gly Arg Pro Val Tyr Asp Gly Gln AlaTrp Val Leu Ala Ala 260 265 270 Val Ala Cys Thr Val Leu Arg Cys Leu GlyIle Pro Ala Arg Val Val 275 280 285 Thr Thr Phe Ala Ser Ala Gln Gly ThrGly Gly Arg Leu Leu Ile Asp 290 295 300 Glu Tyr Tyr Asn Glu Glu Gly LeuGln Asn Gly Glu Gly Gln Arg Gly 305 310 315 320 Arg Ile Trp Ile Phe GlnThr Ser Thr Glu Cys Trp Met Thr Arg Pro 325 330 335 Ala Leu Pro Gln GlyTyr Asp Gly Trp Gln Ile Leu Asp Pro Ser Ala 340 345 350 Pro Asn Gly GlyGly Val Leu Gly Ser Cys Asp Leu Val Pro Val Arg 355 360 365 Ala Val LysGlu Gly Thr Val Gly Leu Thr Pro Ala Val Ser Asp Leu 370 375 380 Phe AlaAla Ile Asn Ala Ser Cys Val Val Trp Lys Cys Cys Glu Asp 385 390 395 400Gly Thr Leu Glu Leu Thr Asp Ser Asn Thr Lys Tyr Val Gly Asn Asn 405 410415 Ile Ser Thr Lys Gly Val Gly Ser Asp Arg Cys Glu Asp Ile Thr Gln 420425 430 Asn Tyr Lys Tyr Pro Glu Gly Ser Leu Gln Glu Lys Glu Val Leu Glu435 440 445 Arg Val Glu Lys Glu Lys Met Glu Arg Glu Lys Asp Asn Gly IleArg 450 455 460 Pro Pro Ser Leu Glu Thr Ala Ser Pro Leu Tyr Leu Leu LeuLys Ala 465 470 475 480 Pro Ser Ser Leu Pro Leu Arg Gly Asp Ala Gln IleSer Val Thr Leu 485 490 495 Val Asn His Ser Glu Gln Glu Lys Ala Val GlnLeu Ala Ile Gly Val 500 505 510 Gln Ala Val His Tyr Asn Gly Val Leu AlaAla Lys Leu Trp Arg Lys 515 520 525 Lys Leu His Leu Thr Leu Ser Ala AsnLeu Glu Lys Ile Ile Thr Ile 530 535 540 Gly Leu Phe Phe Ser Asn Phe GluArg Asn Pro Pro Glu Asn Thr Phe 545 550 555 560 Leu Arg Leu Thr Ala MetAla Thr His Ser Glu Ser Asn Leu Ser Cys 565 570 575 Phe Ala Gln Glu AspIle Ala Ile Cys Arg Pro His Leu Ala Ile Lys 580 585 590 Met Pro Glu LysAla Glu Gln Tyr Gln Pro Leu Thr Ala Ser Val Ser 595 600 605 Leu Gln AsnSer Leu Asp Ala Pro Met Glu Asp Cys Val Ile Ser Ile 610 615 620 Leu GlyArg Gly Leu Ile His Arg Glu Arg Ser Tyr Arg Phe Arg Ser 625 630 635 640Val Trp Pro Glu Asn Thr Met Cys Ala Lys Phe Gln Phe Thr Pro Thr 645 650655 His Val Gly Leu Gln Arg Leu Thr Val Glu Val Asp Cys Asn Met Phe 660665 670 Gln Asn Leu Thr Asn Tyr Lys Ser Val Thr Val Val Ala Pro Glu Leu675 680 685 Ser Ala 690 134 706 PRT Homo sapiens 134 Thr Gln Gly Ile ArgVal Thr Lys Val Asp Trp Gln Arg Ser Arg Asn 1 5 10 15 Gly Ala Ala HisHis Thr Gln Glu Tyr Pro Cys Pro Glu Leu Val Val 20 25 30 Arg Arg Gly GlnSer Phe Ser Leu Thr Leu Glu Leu Ser Arg Ala Leu 35 40 45 Asp Cys Glu GluIle Leu Ile Phe Thr Val Glu Thr Gly Pro Arg Ala 50 55 60 Ser Glu Ala LeuHis Thr Lys Ala Val Phe Gln Thr Ser Glu Leu Glu 65 70 75 80 Arg Gly GluGly Trp Thr Ala Ala Arg Glu Ala Gln Met Glu Lys Thr 85 90 95 Leu Thr ValSer Leu Ala Ser Pro Pro Ser Ala Val Ile Gly Arg Tyr 100 105 110 Leu LeuSer Ile Arg Leu Ser Ser His Arg Lys His Ser Asn Arg Arg 115 120 125 LeuGly Glu Phe Val Leu Leu Phe Asn Pro Trp Cys Ala Glu Asp Asp 130 135 140Val Phe Leu Ala Ser Glu Glu Glu Arg Gln Glu Tyr Val Leu Ser Asp 145 150155 160 Ser Gly Ile Ile Phe Arg Gly Val Glu Lys His Ile Arg Ala Gln Gly165 170 175 Trp Asn Tyr Gly Gln Phe Glu Glu Asp Ile Leu Asn Ile Cys LeuSer 180 185 190 Ile Leu Asp Arg Ser Pro Gly His Gln Asn Asn Pro Ala ThrAsp Val 195 200 205 Ser Cys Arg His Asn Pro Ile Tyr Val Thr Arg Val IleSer Ala Met 210 215 220 Val Asn Ser Asn Asn Asp Arg Gly Val Val Gln GlyGln Trp Gln Gly 225 230 235 240 Lys Tyr Gly Gly Gly Thr Ser Pro Leu HisTrp Arg Gly Ser Val Ala 245 250 255 Ile Leu Gln Lys Trp Leu Lys Gly ArgTyr Lys Pro Val Lys Tyr Gly 260 265 270 Gln Cys Trp Val Phe Ala Gly ValLeu Cys Thr Val Leu Arg Cys Leu 275 280 285 Gly Ile Ala Thr Arg Val ValSer Asn Phe Asn Ser Ala His Asp Thr 290 295 300 Asp Gln Asn Leu Ser ValAsp Lys Tyr Val Asp Ser Phe Gly Arg Thr 305 310 315 320 Leu Glu Asp LeuThr Glu Asp Ser Met Trp Asn Phe His Val Trp Asn 325 330 335 Glu Ser TrpPhe Ala Arg Gln Asp Leu Gly Pro Ser Tyr Asn Gly Trp 340 345 350 Gln ValLeu Asp Ala Thr Pro Gln Glu Glu Ser Glu Gly Val Phe Arg 355 360 365 CysGly Pro Ala Ser Val Thr Ala Ile Arg Glu Gly Asp Val His Leu 370 375 380Ala His Asp Gly Pro Phe Val Phe Ala Glu Val Asn Ala Asp Tyr Ile 385 390395 400 Thr Trp Leu Trp His Glu Asp Glu Ser Arg Glu Arg Val Tyr Ser Asn405 410 415 Thr Lys Lys Ile Gly Arg Cys Ile Ser Thr Lys Ala Val Gly SerAsp 420 425 430 Ser Arg Val Asp Ile Thr Asp Leu Tyr Lys Tyr Pro Glu GlySer Arg 435 440 445 Lys Glu Arg Gln Val Tyr Ser Lys Ala Val Asn Arg LeuPhe Gly Val 450 455 460 Glu Ala Ser Gly Arg Arg Ile Trp Ile Arg Arg AlaGly Gly Arg Cys 465 470 475 480 Leu Trp Arg Asp Asp Leu Leu Glu Pro AlaThr Lys Pro Ser Ile Ala 485 490 495 Gly Lys Phe Lys Val Leu Glu Pro ProMet Leu Gly His Asp Leu Arg 500 505 510 Leu Ala Leu Cys Leu Ala Asn LeuThr Ser Arg Ala Gln Arg Val Arg 515 520 525 Val Asn Leu Ser Gly Ala ThrIle Leu Tyr Thr Arg Lys Pro Val Ala 530 535 540 Glu Ile Leu His Glu SerHis Ala Val Arg Leu Gly Pro Gln Glu Glu 545 550 555 560 Lys Arg Ile ProIle Thr Ile Ser Tyr Ser Lys Tyr Lys Glu Asp Leu 565 570 575 Thr Glu AspLys Lys Ile Leu Leu Ala Ala Met Cys Leu Val Thr Lys 580 585 590 Gly GluLys Leu Leu Val Glu Lys Asp Ile Thr Leu Glu Asp Phe Ile 595 600 605 ThrIle Lys Val Leu Gly Pro Ala Met Val Gly Val Ala Val Thr Val 610 615 620Glu Val Thr Val Val Asn Pro Leu Ile Glu Arg Val Lys Asp Cys Ala 625 630635 640 Leu Met Val Glu Gly Ser Gly Leu Leu Gln Glu Gln Leu Ser Ile Asp645 650 655 Val Pro Thr Leu Glu Pro Gln Glu Arg Ala Ser Val Gln Phe AspIle 660 665 670 Thr Pro Ser Lys Ser Gly Pro Arg Gln Leu Gln Val Asp LeuVal Ser 675 680 685 Pro His Phe Pro Asp Ile Lys Gly Phe Val Ile Val HisVal Ala Thr 690 695 700 Ala Lys 705 135 685 PRT Homo sapiens 135 Ala AlaLeu Gly Val Gln Ser Ile Asn Trp Gln Lys Ala Leu Asn Arg 1 5 10 15 GlnAla His His Thr Asp Lys Phe Ser Ser Gln Glu Leu Ile Leu Arg 20 25 30 ArgGly Gln Asn Phe Gln Val Leu Ile Ile Met Asn Lys Gly Leu Gly 35 40 45 SerAsn Glu Arg Leu Glu Phe Ile Asp Thr Thr Gly Pro Tyr Pro Ser 50 55 60 GluSer Ala Met Thr Lys Ala Val Phe Pro Leu Ser Asn Gly Ser Ser 65 70 75 80Gly Gly Trp Ser Ala Val Leu Gln Ala Ser Asn Gly Asn Thr Leu Thr 85 90 95Ile Ser Ile Ser Ser Pro Ala Ser Ala Pro Ile Gly Arg Tyr Thr Met 100 105110 Ala Leu Gln Ile Phe Ser Gln Gly Gly Ile Ser Ser Val Lys Leu Gly 115120 125 Thr Phe Ile Leu Leu Phe Asn Pro Trp Leu Asn Val Asp Ser Val Phe130 135 140 Met Gly Asn His Ala Glu Arg Glu Glu Tyr Val Gln Glu Asp AlaGly 145 150 155 160 Ile Ile Phe Val Gly Ser Thr Asn Arg Ile Gly Met IleGly Trp Asn 165 170 175 Phe Gly Gln Phe Glu Glu Asp Ile Leu Ser Ile CysLeu Ser Ile Leu 180 185 190 Asp Arg Ser Leu Asn Phe Arg Arg Asp Ala AlaThr Asp Val Ala Ser 195 200 205 Arg Asn Asp Pro Lys Tyr Val Gly Arg ValLeu Ser Ala Met Ile Asn 210 215 220 Ser Asn Asp Asp Asn Gly Val Leu AlaGly Asn Trp Ser Gly Thr Tyr 225 230 235 240 Thr Gly Gly Arg Asp Pro ArgSer Trp Asp Gly Ser Val Glu Ile Leu 245 250 255 Lys Asn Trp Lys Lys SerGly Phe Ser Pro Val Arg Tyr Gly Gln Cys 260 265 270 Trp Val Phe Leu ArgSer Leu Gly Ile Pro Ser Arg Val Ile Thr Asn 275 280 285 Phe Asn Ser AlaHis Asp Thr Asp Arg Asn Leu Ser Val Asp Val Tyr 290 295 300 Tyr Asp ProMet Gly Asn Pro Leu Asp Lys Gly Ser Asp Ser Val Trp 305 310 315 320 AsnPhe His Val Trp Asn Glu Gly Trp Phe Val Arg Ser Asp Leu Gly 325 330 335Pro Pro Tyr Gly Gly Trp Gln Val Leu Asp Ala Thr Pro Gln Glu Arg 340 345350 Ser Gln Gly Val Phe Gln Cys Gly Pro Ala Ser Val Ile Gly Val Arg 355360 365 Glu Gly Asp Val Gln Leu Asn Phe Asp Met Pro Phe Ile Phe Ala Glu370 375 380 Val Asn Ala Asp Arg Ile Thr Trp Leu Tyr Asp Asn Thr Thr GlyLys 385 390 395 400 Gln Trp Lys Asn Ser Val Asn Ser His Thr Ile Gly ArgTyr Ile Ser 405 410 415 Thr Lys Ala Val Gly Ser Asn Ala Arg Met Asp ValThr Asp Lys Tyr 420 425 430 Lys Tyr Pro Glu Gly Ser Asp Gln Glu Arg GlnVal Phe Gln Lys Ala 435 440 445 Leu Gly Lys Leu Lys Pro Asn Thr Pro PheAla Ala Thr Ser Ser Met 450 455 460 Gly Leu Glu Thr Glu Glu Gln Glu ProSer Ile Ser Gly Lys Leu Lys 465 470 475 480 Val Ala Gly Met Leu Ala ValGly Lys Glu Val Asn Leu Val Leu Leu 485 490 495 Leu Lys Asn Leu Ser ArgAsp Thr Lys Thr Val Thr Val Asn Met Thr 500 505 510 Ala Trp Thr Ile IleTyr Asn Gly Thr Leu Val His Glu Val Trp Lys 515 520 525 Asp Ser Ala ThrMet Ser Leu Asp Pro Glu Glu Glu Ala Glu His Pro 530 535 540 Ile Lys IleSer Tyr Ala Gln Tyr Glu Arg Tyr Leu Lys Ser Asp Asn 545 550 555 560 MetIle Arg Ile Thr Ala Val Cys Lys Val Pro Asp Glu Ser Glu Val 565 570 575Val Val Glu Arg Asp Ile Ile Leu Asp Asn Pro Thr Leu Thr Leu Glu 580 585590 Val Leu Asn Glu Ala Arg Val Arg Lys Pro Val Asn Val Gln Met Leu 595600 605 Phe Ser Asn Pro Leu Asp Glu Pro Val Arg Asp Cys Val Leu Met Val610 615 620 Glu Gly Ser Gly Leu Leu Leu Gly Asn Leu Lys Ile Asp Val ProThr 625 630 635 640 Leu Gly Pro Lys Glu Arg Ser Arg Val Arg Phe Asp IleLeu Pro Ser 645 650 655 Arg Ser Gly Thr Lys Gln Leu Leu Ala Asp Phe SerCys Asn Lys Phe 660 665 670 Pro Ala Ile Lys Ala Met Leu Ser Ile Asp ValAla Glu 675 680 685 136 684 PRT Homo sapiens 136 Ala Glu Glu Leu Val LeuGlu Arg Cys Asp Leu Glu Leu Glu Thr Asn 1 5 10 15 Gly Arg Asp His HisThr Ala Asp Leu Cys Arg Glu Lys Leu Val Val 20 25 30 Arg Arg Gly Gln ProPhe Trp Leu Thr Leu His Phe Glu Gly Arg Asn 35 40 45 Tyr Glu Ala Ser ValAsp Ser Leu Thr Phe Ser Val Val Thr Gly Pro 50 55 60 Ala Pro Ser Gln GluAla Gly Thr Lys Ala Arg Phe Pro Leu Arg Asp 65 70 75 80 Ala Val Glu GluGly Asp Trp Thr Ala Thr Val Val Asp Gln Gln Asp 85 90 95 Cys Thr Leu SerLeu Gln Leu Thr Thr Pro Ala Asn Ala Pro Ile Gly 100 105 110 Leu Tyr ArgLeu Ser Leu Glu Ala Ser Thr Gly Tyr Gln Gly Ser Ser 115 120 125 Phe ValLeu Gly His Phe Ile Leu Leu Phe Asn Ala Trp Cys Pro Ala 130 135 140 AspAla Val Tyr Leu Asp Ser Glu Glu Glu Arg Gln Glu Tyr Val Leu 145 150 155160 Thr Gln Gln Gly Phe Ile Tyr Gln Gly Ser Ala Lys Phe Ile Lys Asn 165170 175 Ile Pro Trp Asn Phe Gly Gln Phe Gln Asp Gly Ile Leu Asp Ile Cys180 185 190 Leu Ile Leu Leu Asp Val Asn Pro Lys Phe Leu Lys Asn Ala GlyArg 195 200 205 Asp Cys Ser Arg Arg Ser Ser Pro Val Tyr Val Gly Arg ValGly Ser 210 215 220 Gly Met Val Asn Cys Asn Asp Asp Gln Gly Val Leu LeuGly Arg Trp 225 230 235 240 Asp Asn Asn Tyr Gly Asp Gly Val Ser Pro MetSer Trp Ile Gly Ser 245 250 255 Val Asp Ile Leu Arg Arg Trp Lys Asn HisGly Cys Gln Arg Val Lys 260 265 270 Tyr Gly Gln Cys Trp Val Phe Ala AlaVal Ala Cys Thr Val Leu Arg 275 280 285 Cys Leu Gly Ile Pro Thr Arg ValVal Thr Asn Tyr Asn Ser Ala His 290 295 300 Asp Gln Asn Ser Asn Leu LeuIle Glu Tyr Phe Arg Asn Glu Phe Gly 305 310 315 320 Glu Ile Gln Gly AspLys Ser Glu Met Ile Trp Asn Phe His Cys Trp 325 330 335 Val Glu Ser TrpMet Thr Arg Pro Asp Leu Gln Pro Gly Tyr Glu Gly 340 345 350 Trp Gln AlaLeu Asp Pro Thr Pro Gln Glu Lys Ser Glu Gly Thr Tyr 355 360 365 Cys CysGly Pro Val Pro Val Arg Ala Ile Lys Glu Gly Asp Leu Ser 370 375 380 ThrLys Tyr Asp Ala Pro Phe Val Phe Ala Glu Val Asn Ala Asp Val 385 390 395400 Val Asp Trp Ile Gln Gln Asp Asp Gly Ser Val His Lys Ser Ile Asn 405410 415 Arg Ser Leu Ile Val Gly Leu Lys Ile Ser Thr Lys Ser Val Gly Arg420 425 430 Asp Glu Arg Glu Asp Ile Thr His Thr Tyr Lys Tyr Pro Glu GlySer 435 440 445 Ser Glu Glu Arg Glu Ala Phe Thr Arg Ala Asn His Leu AsnLys Leu 450 455 460 Ala Glu Lys Glu Glu Thr Gly Met Ala Met Arg Ile ArgVal Gly Gln 465 470 475 480 Ser Met Asn Met Gly Ser Asp Phe Asp Val PheAla His Ile Thr Asn 485 490 495 Asn Thr Ala Glu Glu Tyr Val Cys Arg LeuLeu Leu Cys Ala Arg Thr 500 505 510 Val Ser Tyr Asn Gly Ile Leu Gly ProGlu Cys Gly Thr Lys Tyr Leu 515 520 525 Leu Asn Leu Thr Leu Glu Pro PheSer Glu Lys Ser Val Pro Leu Cys 530 535 540 Ile Leu Tyr Glu Lys Tyr ArgAsp Cys Leu Thr Glu Ser Asn Leu Ile 545 550 555 560 Lys Val Arg Ala LeuLeu Val Glu Pro Val Ile Asn Ser Tyr Leu Leu 565 570 575 Ala Glu Arg AspLeu Tyr Asn Pro Glu Ile Lys Ile Arg Ile Leu Gly 580 585 590 Glu Pro LysGln Lys Arg Lys Leu Val Ala Glu Val Ser Leu Gln Asn 595 600 605 Pro LeuPro Val Ala Leu Glu Gly Cys Thr Phe Thr Val Glu Gly Ala 610 615 620 GlyLeu Thr Glu Glu Gln Lys Thr Val Glu Ile Pro Asp Pro Val Glu 625 630 635640 Ala Gly Glu Glu Val Lys Val Arg Met Asp Leu Val Pro Leu His Met 645650 655 Gly Leu His Lys Leu Val Val Asn Phe Glu Ser Asp Lys Leu Lys Ala660 665 670 Val Lys Gly Phe Arg Asn Val Ile Ile Gly Pro Ala 675 680 137618 PRT Homo sapiens 137 Ser Glu Thr Ser Arg Thr Ala Phe Gly Gly Arg ArgAla Val Pro Pro 1 5 10 15 Asn Asn Ser Asn Ala Ala Glu Asp Asp Leu ProThr Val Glu Leu Gln 20 25 30 Gly Val Val Pro Arg Gly Val Asn Leu Gln GluPhe Leu Asn Val Thr 35 40 45 Ser Val His Leu Phe Lys Glu Arg Trp Asp ThrAsn Lys Val Asp His 50 55 60 His Thr Asp Lys Tyr Glu Asn Asn Lys Leu IleVal Arg Arg Gly Gln 65 70 75 80 Ser Phe Tyr Val Gln Ile Asp Phe Ser ArgPro Tyr Asp Pro Arg Arg 85 90 95 Asp Leu Phe Arg Val Glu Tyr Val Ile GlyArg Tyr Pro Gln Glu Asn 100 105 110 Lys Gly Thr Tyr Ile Pro Val Pro IleVal Ser Glu Leu Gln Ser Gly 115 120 125 Lys Trp Gly Ala Lys Ile Val MetArg Glu Asp Arg Ser Val Arg Leu 130 135 140 Ser Ile Gln Ser Ser Pro LysCys Ile Val Gly Lys Phe Arg Met Tyr 145 150 155 160 Val Ala Val Trp ThrPro Tyr Gly Val Leu Arg Thr Ser Arg Asn Pro 165 170 175 Glu Thr Asp ThrTyr Ile Leu Phe Asn Pro Trp Cys Glu Asp Asp Ala 180 185 190 Val Tyr LeuAsp Asn Glu Lys Glu Arg Glu Glu Tyr Val Leu Asn Asp 195 200 205 Ile GlyVal Ile Phe Tyr Gly Glu Val Asn Asp Ile Lys Thr Arg Ser 210 215 220 TrpSer Tyr Gly Gln Phe Glu Asp Gly Ile Leu Asp Thr Cys Leu Tyr 225 230 235240 Val Met Asp Arg Ala Gln Met Asp Leu Ser Gly Arg Gly Asn Pro Ile 245250 255 Lys Val Ser Arg Val Gly Ser Ala Met Val Asn Ala Lys Asp Asp Glu260 265 270 Gly Val Leu Val Gly Ser Trp Asp Asn Ile Tyr Ala Tyr Gly ValPro 275 280 285 Pro Ser Ala Trp Thr Gly Ser Val Asp Ile Leu Leu Glu TyrArg Ser 290 295 300 Ser Glu Asn Pro Val Arg Tyr Gly Gln Cys Trp Val PheAla Gly Val 305 310 315 320 Phe Asn Thr Phe Leu Arg Cys Leu Gly Ile ProAla Arg Ile Val Thr 325 330 335 Asn Tyr Phe Ser Ala His Asp Asn Asp AlaAsn Leu Gln Met Asp Ile 340 345 350 Phe Leu Glu Glu Asp Gly Asn Val AsnSer Lys Leu Thr Lys Asp Ser 355 360 365 Val Trp Asn Tyr His Cys Trp AsnGlu Ala Trp Met Thr Arg Pro Asp 370 375 380 Leu Pro Val Gly Phe Gly GlyTrp Gln Ala Val Asp Ser Thr Pro Gln 385 390 395 400 Glu Asn Ser Asp GlyMet Tyr Arg Cys Gly Pro Ala Ser Val Gln Ala 405 410 415 Ile Lys His GlyHis Val Cys Phe Gln Phe Asp Ala Pro Phe Val Phe 420 425 430 Ala Glu ValAsn Ser Asp Leu Ile Tyr Ile Thr Ala Lys Lys Asp Gly 435 440 445 Thr HisVal Val Glu Asn Val Asp Ala Thr His Ile Gly Lys Leu Ile 450 455 460 ValThr Lys Gln Ile Gly Gly Asp Gly Met Met Asp Ile Thr Asp Thr 465 470 475480 Tyr Lys Phe Gln Glu Gly Gln Glu Glu Glu Arg Leu Ala Leu Glu Thr 485490 495 Ala Leu Met Tyr Gly Ala Lys Lys Pro Leu Asn Thr Glu Gly Val Met500 505 510 Lys Ser Arg Ile Pro Glu Ile Ile Ile Lys Val Arg Gly Thr GlnVal 515 520 525 Val Gly Ser Asp Met Thr Val Thr Val Gln Phe Thr Asn ProLeu Lys 530 535 540 Glu Thr Leu Arg Asn Val Trp Val His Leu Asp Gly ProGly Val Thr 545 550 555 560 Arg Pro Met Lys Lys Met Phe Arg Glu Ile ArgPro Asn Ser Thr Val 565 570 575 Gln Trp Glu Glu Val Cys Arg Pro Trp ValSer Gly His Arg Lys Leu 580 585 590 Ile Ala Ser Met Ser Ser Asp Ser LeuArg His Val Tyr Gly Glu Leu 595 600 605 Asp Val Gln Ile Gln Arg Arg ProSer Met 610 615 138 812 PRT Homo sapiens 138 Met Asp Gly Pro Arg Ser AspVal Gly Arg Trp Gly Gly Asn Pro Leu 1 5 10 15 Gln Pro Pro Thr Thr ProSer Pro Glu Pro Glu Pro Asp Gly Arg Ser 20 25 30 Arg Arg Gly Gly Gly ArgSer Phe Trp Ala Arg Cys Cys Gly Cys Cys 35 40 45 Ser Cys Arg Asn Ala AlaAsp Asp Asp Trp Gly Pro Glu Pro Ser Asp 50 55 60 Ser Arg Gly Arg Gly SerSer Ser Gly Thr Arg Arg Pro Gly Ser Arg 65 70 75 80 Gly Ser Asp Ser ArgArg Pro Val Ser Arg Gly Ser Gly Val Asn Ala 85 90 95 Ala Gly Asp Gly ThrIle Arg Glu Gly Met Leu Val Val Asn Gly Val 100 105 110 Asp Leu Leu SerSer Arg Ser Asp Gln Asn Arg Arg Glu His His Thr 115 120 125 Asp Glu TyrGlu Tyr Asp Glu Leu Ile Val Arg Arg Gly Gln Pro Phe 130 135 140 His MetLeu Leu Leu Leu Ser Arg Thr Tyr Glu Ser Ser Asp Arg Ile 145 150 155 160Thr Leu Glu Leu Leu Ile Gly Asn Asn Pro Glu Val Gly Lys Gly Thr 165 170175 His Val Ile Ile Pro Val Gly Lys Gly Gly Ser Gly Gly Trp Lys Ala 180185 190 Gln Val Val Lys Ala Ser Gly Gln Asn Leu Asn Leu Arg Val His Thr195 200 205 Ser Pro Asn Ala Ile Ile Gly Lys Phe Gln Phe Thr Val Arg ThrGln 210 215 220 Ser Asp Ala Gly Glu Phe Gln Leu Pro Phe Asp Pro Arg AsnGlu Ile 225 230 235 240 Tyr Ile Leu Phe Asn Pro Trp Cys Pro Glu Asp IleVal Tyr Val Asp 245 250 255 His Glu Asp Trp Arg Gln Glu Tyr Val Leu AsnGlu Ser Gly Arg Ile 260 265 270 Tyr Tyr Gly Thr Glu Ala Gln Ile Gly GluArg Thr Trp Asn Tyr Gly 275 280 285 Gln Phe Asp His Gly Val Leu Asp AlaCys Leu Tyr Ile Leu Asp Arg 290 295 300 Arg Gly Met Pro Tyr Gly Gly ArgGly Asp Pro Val Asn Val Ser Arg 305 310 315 320 Val Ile Ser Ala Met ValAsn Ser Leu Asp Asp Asn Gly Val Leu Ile 325 330 335 Gly Asn Trp Ser GlyAsp Tyr Ser Arg Gly Thr Asn Pro Ser Ala Trp 340 345 350 Val Gly Ser ValGlu Ile Leu Leu Ser Tyr Leu Arg Thr Gly Tyr Ser 355 360 365 Val Pro TyrGly Gln Cys Trp Val Phe Ala Gly Val Thr Thr Thr Val 370 375 380 Leu ArgCys Leu Gly Leu Ala Thr Arg Thr Val Thr Asn Phe Asn Ser 385 390 395 400Ala His Asp Thr Asp Thr Ser Leu Thr Met Asp Ile Tyr Phe Asp Glu 405 410415 Asn Met Lys Pro Leu Glu His Leu Asn His Asp Ser Val Trp Asn Phe 420425 430 His Val Trp Asn Asp Cys Trp Met Lys Arg Pro Asp Leu Pro Ser Gly435 440 445 Phe Asp Gly Trp Gln Val Val Asp Ala Thr Pro Gln Glu Thr SerSer 450 455 460 Gly Ile Phe Cys Cys Gly Pro Cys Ser Val Glu Ser Ile LysAsn Gly 465 470 475 480 Leu Val Tyr Met Lys Tyr Asp Thr Pro Phe Ile PheAla Glu Val Asn 485 490 495 Ser Asp Lys Val Tyr Trp Gln Arg Gln Asp AspGly Ser Phe Lys Ile 500 505 510 Val Tyr Val Glu Glu Lys Ala Ile Gly ThrLeu Ile Val Thr Lys Ala 515 520 525 Ile Ser Ser Asn Met Arg Glu Asp IleThr Tyr Leu Tyr Lys His Pro 530 535 540 Glu Gly Ser Asp Ala Glu Arg LysAla Val Glu Thr Ala Ala Ala His 545 550 555 560 Gly Ser Lys Pro Asn ValTyr Ala Asn Arg Gly Ser Glu Asp Val Ala 565 570 575 Met Gln Val Glu AlaGln Asp Ala Val Met Gly Gln Asp Leu Met Val 580 585 590 Ser Val Met LeuIle Asn His Ser Ser Ser Arg Arg Thr Val Lys Leu 595 600 605 His Leu TyrLeu Ser Val Thr Phe Tyr Thr Gly Val Ser Gly Thr Ile 610 615 620 Phe LysGlu Thr Lys Lys Glu Val Glu Leu Ala Pro Gly Ala Ser Asp 625 630 635 640Arg Val Thr Met Pro Val Ala Tyr Lys Glu Tyr Arg Pro His Leu Val 645 650655 Asp Gln Gly Ala Met Leu Leu Asn Val Ser Gly His Val Lys Glu Ser 660665 670 Gly Gln Val Leu Ala Lys Gln His Thr Phe Arg Leu Thr Pro Asp Leu675 680 685 Ser Leu Thr Leu Leu Gly Ala Ala Val Val Gly Gln Glu Cys GluVal 690 695 700 Gln Ile Val Phe Lys Asn Pro Leu Pro Val Thr Leu Thr AsnVal Val 705 710 715 720 Phe Arg Leu Glu Gly Ser Gly Leu Gln Arg Pro LysIle Leu Asn Val 725 730 735 Gly Asp Ile Gly Gly Asn Glu Thr Val Thr LeuArg Gln Ser Phe Val 740 745 750 Pro Val Arg Pro Gly Pro Arg Gln Leu IleAla Ser Leu Asp Ser Pro 755 760 765 Gln Leu Ser Gln Val His Gly Val IleGln Val Asp Val Ala Pro Ala 770 775 780 Pro Gly Asp Gly Gly Phe Phe SerAsp Ala Gly Gly Asp Ser His Leu 785 790 795 800 Gly Glu Thr Ile Pro MetAla Ser Arg Gly Gly Ala 805 810 139 683 PRT Homo sapiens 139 Met Asp AlaSer Lys Glu Leu Gln Val Leu His Ile Asp Phe Leu Asn 1 5 10 15 Gln AspAsn Ala Val Ser His His Thr Trp Glu Phe Gln Thr Ser Ser 20 25 30 Pro ValPhe Arg Arg Gly Gln Val Phe His Leu Arg Leu Val Leu Asn 35 40 45 Gln ProLeu Gln Ser Tyr His Gln Leu Lys Leu Glu Phe Ser Thr Gly 50 55 60 Pro AsnPro Ser Ile Ala Lys His Thr Leu Val Val Leu Asp Pro Arg 65 70 75 80 ThrPro Ser Asp His Tyr Asn Trp Gln Ala Thr Leu Gln Asn Glu Ser 85 90 95 GlyLys Glu Val Thr Val Ala Val Thr Ser Ser Pro Asn Ala Ile Leu 100 105 110Gly Lys Tyr Gln Leu Asn Val Lys Thr Gly Asn His Ile Leu Lys Ser 115 120125 Glu Glu Asn Ile Leu Tyr Leu Leu Phe Asn Pro Trp Cys Lys Glu Asp 130135 140 Met Val Phe Met Pro Asp Glu Asp Glu Arg Lys Glu Tyr Ile Leu Asn145 150 155 160 Asp Thr Gly Cys His Tyr Val Gly Ala Ala Arg Ser Ile LysCys Lys 165 170 175 Pro Trp Asn Phe Gly Gln Phe Glu Lys Asn Val Leu AspCys Cys Ile 180 185 190 Ser Leu Leu Thr Glu Ser Ser Leu Lys Pro Thr AspArg Arg Asp Pro 195 200 205 Val Leu Val Cys Arg Ala Met Cys Ala Met MetSer Phe Glu Lys Gly 210 215 220 Gln Gly Val Leu Ile Gly Asn Trp Thr GlyAsp Tyr Glu Gly Gly Thr 225 230 235 240 Ala Pro Tyr Lys Trp Thr Gly SerAla Pro Ile Leu Gln Gln Tyr Tyr 245 250 255 Asn Thr Lys Gln Ala Val CysPhe Gly Gln Cys Trp Val Phe Ala Gly 260 265 270 Ile Leu Thr Thr Val LeuArg Ala Leu Gly Ile Pro Ala Arg Ser Val 275 280 285 Thr Gly Phe Asp SerAla His Asp Thr Glu Arg Asn Leu Thr Val Asp 290 295 300 Thr Tyr Val AsnGlu Asn Gly Lys Lys Ile Thr Ser Met Thr His Asp 305 310 315 320 Ser ValTrp Asn Phe His Val Trp Thr Asp Ala Trp Met Lys Arg Pro 325 330 335 AspLeu Pro Lys Gly Tyr Asp Gly Trp Gln Ala Val Asp Ala Thr Pro 340 345 350Gln Glu Arg Ser Gln Gly Val Phe Cys Cys Gly Pro Ser Pro Leu Thr 355 360365 Ala Ile Arg Lys Gly Asp Ile Phe Ile Val Tyr Asp Thr Arg Phe Val 370375 380 Phe Ser Glu Val Asn Gly Asp Arg Leu Ile Trp Leu Val Lys Met Val385 390 395 400 Asn Gly Gln Glu Glu Leu His Val Ile Ser Met Glu Thr ThrSer Ile 405 410 415 Gly Lys Asn Ile Ser Thr Lys Ala Val Gly Gln Asp ArgArg Arg Asp 420 425 430 Ile Thr Tyr Glu Tyr Lys Tyr Pro Glu Gly Ser SerGlu Glu Arg Gln 435 440 445 Val Met Asp His Ala Phe Leu Leu Leu Ser SerGlu Arg Glu His Arg 450 455 460 Arg Pro Val Lys Glu Asn Phe Leu His MetSer Val Gln Ser Asp Asp 465 470 475 480 Val Leu Leu Gly Asn Ser Val AsnPhe Thr Val Ile Leu Lys Arg Lys 485 490 495 Thr Ala Ala Leu Gln Asn ValAsn Ile Leu Gly Ser Phe Glu Leu Gln 500 505 510 Leu Tyr Thr Gly Lys LysMet Ala Lys Leu Cys Asp Leu Asn Lys Thr 515 520 525 Ser Gln Ile Gln GlyGln Val Ser Glu Val Thr Leu Thr Leu Asp Ser 530 535 540 Lys Thr Tyr IleAsn Ser Leu Ala Ile Leu Asp Asp Glu Pro Val Ile 545 550 555 560 Arg GlyPhe Ile Ile Ala Glu Ile Val Glu Ser Lys Glu Ile Met Ala 565 570 575 SerGlu Val Phe Thr Ser Phe Gln Tyr Pro Glu Phe Ser Ile Glu Leu 580 585 590Pro Asn Thr Gly Arg Ile Gly Gln Leu Leu Val Cys Asn Cys Ile Phe 595 600605 Lys Asn Thr Leu Ala Ile Pro Leu Thr Asp Val Lys Phe Ser Leu Glu 610615 620 Ser Leu Gly Ile Ser Ser Leu Gln Thr Ser Asp His Gly Thr Val Gln625 630 635 640 Pro Gly Glu Thr Ile Gln Ser Gln Ile Lys Cys Thr Pro IleLys Thr 645 650 655 Gly Pro Lys Lys Phe Ile Val Lys Leu Ser Ser Lys GlnVal Lys Glu 660 665 670 Ile Asn Ala Gln Lys Ile Val Leu Ile Thr Lys 675680 140 709 PRT Homo sapiens 140 Asp Gln Val Ala Thr Leu Arg Leu Glu SerVal Asp Leu Gln Ser Ser 1 5 10 15 Arg Asn Asn Lys Glu His His Thr GlnGlu Met Gly Val Lys Arg Leu 20 25 30 Thr Val Arg Arg Gly Gln Pro Phe TyrLeu Arg Leu Ser Phe Ser Arg 35 40 45 Pro Phe Gln Ser Gln Asn Asp His IleThr Phe Val Ala Glu Thr Gly 50 55 60 Pro Lys Pro Ser Glu Leu Leu Gly ThrArg Ala Thr Phe Phe Leu Thr 65 70 75 80 Arg Val Gln Pro Gly Asn Val TrpSer Ala Ser Asp Phe Thr Ile Asp 85 90 95 Ser Asn Ser Leu Gln Val Ser LeuPhe Thr Pro Ala Asn Ala Val Ile 100 105 110 Gly His Tyr Thr Leu Lys IleGlu Ile Ser Gln Gly Gln Gly His Ser 115 120 125 Val Thr Tyr Pro Leu GlyThr Phe Ile Leu Leu Phe Asn Pro Trp Ser 130 135 140 Pro Glu Asp Asp ValTyr Leu Pro Ser Glu Ile Leu Leu Gln Glu Tyr 145 150 155 160 Ile Met ArgAsp Tyr Gly Phe Val Tyr Lys Gly His Glu Arg Phe Ile 165 170 175 Thr SerTrp Pro Trp Asn Tyr Gly Gln Phe Glu Glu Asp Ile Ile Asp 180 185 190 IleCys Phe Glu Ile Leu Asn Lys Ser Leu Tyr His Leu Lys Asn Pro 195 200 205Ala Lys Asp Cys Ser Gln Arg Asn Asp Val Val Tyr Val Cys Arg Val 210 215220 Val Ser Ala Met Ile Asn Ser Asn Asp Asp Asn Gly Val Leu Gln Gly 225230 235 240 Asn Trp Gly Glu Asp Tyr Ser Lys Gly Val Ser Pro Leu Glu TrpLys 245 250 255 Gly Ser Val Ala Ile Leu Gln Gln Trp Ser Ala Arg Gly GlyGln Pro 260 265 270 Val Lys Tyr Gly Gln Cys Trp Val Phe Ala Ser Val MetCys Thr Val 275 280 285 Met Arg Cys Leu Gly Val Pro Thr Arg Val Val SerAsn Phe Arg Ser 290 295 300 Ala His Asn Val Asp Arg Asn Leu Thr Ile AspThr Tyr Tyr Asp Arg 305 310 315 320 Asn Ala Glu Met Leu Ser Thr Gln LysArg Asp Lys Ile Trp Asn Phe 325 330 335 His Val Trp Asn Glu Cys Trp MetIle Arg Lys Asp Leu Pro Pro Gly 340 345 350 Tyr Asn Gly Trp Gln Val LeuAsp Pro Thr Pro Gln Gln Thr Ser Ser 355 360 365 Gly Leu Phe Cys Cys GlyPro Ala Ser Val Lys Ala Ile Arg Glu Gly 370 375 380 Asp Val His Leu AlaTyr Asp Thr Pro Phe Val Tyr Ala Glu Val Asn 385 390 395 400 Ala Asp GluVal Ile Trp Leu Leu Gly Asp Gly Gln Ala Gln Glu Ile 405 410 415 Leu AlaHis Asn Thr Ser Ser Ile Gly Lys Glu Ile Ser Thr Lys Met 420 425 430 ValGly Ser Asp Gln Arg Gln Ser Ile Thr Ser Ser Tyr Lys Tyr Pro 435 440 445Glu Gly Ser Pro Glu Glu Arg Ala Val Phe Met Lys Ala Ser Arg Lys 450 455460 Met Leu Gly Pro Gln Arg Ala Ser Leu Pro Phe Leu Asp Leu Leu Glu 465470 475 480 Ser Gly Gly Leu Arg Asp Gln Pro Ala Gln Leu Gln Leu His LeuAla 485 490 495 Arg Ile Pro Glu Trp Gly Gln Asp Leu Gln Leu Leu Leu ArgIle Gln 500 505 510 Arg Val Pro Asp Ser Thr His Pro Arg Gly Pro Ile GlyLeu Val Val 515 520 525 Arg Phe Cys Ala Gln Ala Leu Leu His Gly Gly GlyThr Gln Lys Pro 530 535 540 Phe Trp Arg His Thr Val Arg Met Asn Leu AspPhe Gly Lys Glu Thr 545 550 555 560 Gln Trp Pro Leu Leu Leu Pro Tyr SerAsn Tyr Arg Asn Lys Leu Thr 565 570 575 Asp Glu Lys Leu Ile Arg Val SerGly Ile Ala Glu Val Glu Glu Thr 580 585 590 Gly Arg Ser Met Leu Val LeuLys Asp Ile Cys Leu Glu Pro Pro His 595 600 605 Leu Ser Ile Glu Val SerGlu Arg Ala Glu Val Gly Lys Ala Leu Arg 610 615 620 Val His Val Thr LeuThr Asn Thr Leu Met Val Ala Leu Ser Ser Cys 625 630 635 640 Thr Met ValLeu Glu Gly Ser Gly Leu Ile Asn Gly Gln Ile Ala Lys 645 650 655 Asp LeuGly Thr Leu Val Ala Gly His Thr Leu Gln Ile Gln Leu Asp 660 665 670 LeuTyr Pro Thr Lys Ala Gly Pro Arg Gln Leu Gln Val Leu Ile Ser 675 680 685Ser Asn Glu Val Lys Glu Ile Lys Gly Tyr Lys Asp Ile Phe Val Thr 690 695700 Val Ala Gly Ala Pro 705 141 2239 DNA Homo sapiens 141 aacccatgacccaggggatc agagtcacca aggtggactg gcagcggtcg aggaatggcg 60 ctgcccaccacacccaggag tacccctgcc ctgagctggt ggttcgcagg ggccagtcgt 120 tcagcctcacgctggagctg agcagagccc tggactgtga ggagatcctc atcttcacgg 180 tggagacaggaccccgggct tctgaggccc tccacaccaa agctgtgttc cagacatcgg 240 agctggagcggggtgagggc tggacagcag caagggaggc tcagatggag aaaactctga 300 ccgtcagtctcgccagccct cccagtgctg tcattggccg ctacctgctg agcatcaggc 360 tttcctctcaccgcaaacac agcaaccgga ggctgggcga gtttgttctc cttttcaacc 420 catggtgtgcagaggacgat gtgtttctgg cctcagagga ggagagacag gagtacgtgc 480 tcagcgacagcggcatcatc ttccgaggcg tggagaagca catacgagcc cagggctgga 540 actacgggcagtttgaggag gacatcctga acatctgcct ctccatcctg gatcgaagcc 600 ccggtcaccaaaacaaccca gccaccgacg tgtcctgccg ccacaacccc atctacgtca 660 ccagggtcatcagtgccatg gtgaacagca acaacgaccg aggtgtggtg caaggacagt 720 ggcagggcaagtacggcggc ggcaccagcc cgctgcactg gcgcggcagc gtggccattc 780 tgcagaagtggctcaagggc aggtacaagc cagtcaagta cggccagtgc tgggtcttcg 840 ccggagtcctgtgcacagtc ctcaggtgct tggggatagc cacacgggtc gtgtccaact 900 tcaactcagcccacgacaca gaccagaacc tgagtgtgga caaatacgtg gactccttcg 960 ggcggaccctggaggacctg acagaagaca gcatgtggaa tttccatgtc tggaatgaga 1020 gctggtttgcccggcaggac ctaggcccct cttacaatgg ctggcaggtt ctggatgcca 1080 ccccccaggaggagagtgaa ggtgtgttcc ggtgcggccc agcctcagtc accgccatcc 1140 gcgagggtgatgtgcacctg gctcacgatg gccccttcgt gtttgcggag gtcaacgccg 1200 actacatcacctggctgtgg cacgaggatg agagccggga gcgtgtatac tcaaacacga 1260 agaagattgggagatgcatc agcaccaagg cggtgggcag tgactcccgc gtggacatca 1320 ctgacctctacaagtatccg gaagggtccc ggaaagagag gcaggtgtac agcaaggcgg 1380 tgaacaggctgttcggcgtg gaagcctctg gaaggagaat ctggatccgc agggctgggg 1440 gtcgctgtctctggcgtgac gacctcctgg agcctgccac caagcccagc atcgctggca 1500 agttcaaggtgctagagcct cccatgctgg gccacgacct gagactggcc ctgtgcttgg 1560 ccaacctcacctcccgggcc cagcgggtga gggtcaacct gagcggtgcc accatcctct 1620 atacccgcaagccagtggca gagatcctgc atgaatccca cgccgtgagg ctggggccgc 1680 aagaagagaagagaatccca attacaatat cttactctaa gtataaagaa gacctgacag 1740 aggacaagaagatcctgttg gctgccatgt gccttgtcac caaaggagag aagcttctgg 1800 tggagaaggacattactcta gaggacttca tcaccatcaa ggttctgggc ccagccatgg 1860 tgggagtggcagttacagtg gaagtgacag tagtcaaccc cctcatagag agagtgaagg 1920 actgtgcgctgatggtggag ggcagcggcc ttctccagga acagctcagc atcgacgtgc 1980 ctaccctggagcctcaggag agggcctcag tccagtttga catcaccccc tccaaaagtg 2040 gcccaaggcagctgcaggtg gaccttgtaa gccctcactt cccggacatc aagggctttg 2100 tgatcgtccatgtggccact gccaagtgat ggatcatgag ggactgagag gggtggattt 2160 ggcccctgtcctcctcctgc ccattctttg tctcttccac atgggagcca ggaggcctca 2220 gttaatcctgcctcaacct 2239 142 707 PRT Homo sapiens 142 Met Thr Gln Gly Ile Arg ValThr Lys Val Asp Trp Gln Arg Ser Arg 1 5 10 15 Asn Gly Ala Ala His HisThr Gln Glu Tyr Pro Cys Pro Glu Leu Val 20 25 30 Val Arg Arg Gly Gln SerPhe Ser Leu Thr Leu Glu Leu Ser Arg Ala 35 40 45 Leu Asp Cys Glu Glu IleLeu Ile Phe Thr Val Glu Thr Gly Pro Arg 50 55 60 Ala Ser Glu Ala Leu HisThr Lys Ala Val Phe Gln Thr Ser Glu Leu 65 70 75 80 Glu Arg Gly Glu GlyTrp Thr Ala Ala Arg Glu Ala Gln Met Glu Lys 85 90 95 Thr Leu Thr Val SerLeu Ala Ser Pro Pro Ser Ala Val Ile Gly Arg 100 105 110 Tyr Leu Leu SerIle Arg Leu Ser Ser His Arg Lys His Ser Asn Arg 115 120 125 Arg Leu GlyGlu Phe Val Leu Leu Phe Asn Pro Trp Cys Ala Glu Asp 130 135 140 Asp ValPhe Leu Ala Ser Glu Glu Glu Arg Gln Glu Tyr Val Leu Ser 145 150 155 160Asp Ser Gly Ile Ile Phe Arg Gly Val Glu Lys His Ile Arg Ala Gln 165 170175 Gly Trp Asn Tyr Gly Gln Phe Glu Glu Asp Ile Leu Asn Ile Cys Leu 180185 190 Ser Ile Leu Asp Arg Ser Pro Gly His Gln Asn Asn Pro Ala Thr Asp195 200 205 Val Ser Cys Arg His Asn Pro Ile Tyr Val Thr Arg Val Ile SerAla 210 215 220 Met Val Asn Ser Asn Asn Asp Arg Gly Val Val Gln Gly GlnTrp Gln 225 230 235 240 Gly Lys Tyr Gly Gly Gly Thr Ser Pro Leu His TrpArg Gly Ser Val 245 250 255 Ala Ile Leu Gln Lys Trp Leu Lys Gly Arg TyrLys Pro Val Lys Tyr 260 265 270 Gly Gln Cys Trp Val Phe Ala Gly Val LeuCys Thr Val Leu Arg Cys 275 280 285 Leu Gly Ile Ala Thr Arg Val Val SerAsn Phe Asn Ser Ala His Asp 290 295 300 Thr Asp Gln Asn Leu Ser Val AspLys Tyr Val Asp Ser Phe Gly Arg 305 310 315 320 Thr Leu Glu Asp Leu ThrGlu Asp Ser Met Trp Asn Phe His Val Trp 325 330 335 Asn Glu Ser Trp PheAla Arg Gln Asp Leu Gly Pro Ser Tyr Asn Gly 340 345 350 Trp Gln Val LeuAsp Ala Thr Pro Gln Glu Glu Ser Glu Gly Val Phe 355 360 365 Arg Cys GlyPro Ala Ser Val Thr Ala Ile Arg Glu Gly Asp Val His 370 375 380 Leu AlaHis Asp Gly Pro Phe Val Phe Ala Glu Val Asn Ala Asp Tyr 385 390 395 400Ile Thr Trp Leu Trp His Glu Asp Glu Ser Arg Glu Arg Val Tyr Ser 405 410415 Asn Thr Lys Lys Ile Gly Arg Cys Ile Ser Thr Lys Ala Val Gly Ser 420425 430 Asp Ser Arg Val Asp Ile Thr Asp Leu Tyr Lys Tyr Pro Glu Gly Ser435 440 445 Arg Lys Glu Arg Gln Val Tyr Ser Lys Ala Val Asn Arg Leu PheGly 450 455 460 Val Glu Ala Ser Gly Arg Arg Ile Trp Ile Arg Arg Ala GlyGly Arg 465 470 475 480 Cys Leu Trp Arg Asp Asp Leu Leu Glu Pro Ala ThrLys Pro Ser Ile 485 490 495 Ala Gly Lys Phe Lys Val Leu Glu Pro Pro MetLeu Gly His Asp Leu 500 505 510 Arg Leu Ala Leu Cys Leu Ala Asn Leu ThrSer Arg Ala Gln Arg Val 515 520 525 Arg Val Asn Leu Ser Gly Ala Thr IleLeu Tyr Thr Arg Lys Pro Val 530 535 540 Ala Glu Ile Leu His Glu Ser HisAla Val Arg Leu Gly Pro Gln Glu 545 550 555 560 Glu Lys Arg Ile Pro IleThr Ile Ser Tyr Ser Lys Tyr Lys Glu Asp 565 570 575 Leu Thr Glu Asp LysLys Ile Leu Leu Ala Ala Met Cys Leu Val Thr 580 585 590 Lys Gly Glu LysLeu Leu Val Glu Lys Asp Ile Thr Leu Glu Asp Phe 595 600 605 Ile Thr IleLys Val Leu Gly Pro Ala Met Val Gly Val Ala Val Thr 610 615 620 Val GluVal Thr Val Val Asn Pro Leu Ile Glu Arg Val Lys Asp Cys 625 630 635 640Ala Leu Met Val Glu Gly Ser Gly Leu Leu Gln Glu Gln Leu Ser Ile 645 650655 Asp Val Pro Thr Leu Glu Pro Gln Glu Arg Ala Ser Val Gln Phe Asp 660665 670 Ile Thr Pro Ser Lys Ser Gly Pro Arg Gln Leu Gln Val Asp Leu Val675 680 685 Ser Pro His Phe Pro Asp Ile Lys Gly Phe Val Ile Val His ValAla 690 695 700 Thr Ala Lys 705 143 2105 DNA Homo sapiens 143 aacccatgacccaggggatc agagtcacca aggtggactg gcagcggtcg aggaatggcg 60 ctgcccaccacacccaggag tacccctgcc ctgagctggt ggttcgcagg ggccagtcgt 120 tcagcctcacgctggagctg agcagagccc tggactgtga ggagatcctc atcttcacgg 180 tggagacaggaccccgggct tctgaggccc tccacaccaa agctgtgttc cagacatcgg 240 agctggagcggggtgagggc tggacagcag caagggaggc tcagatggag aaaactctga 300 ccgtcagtctcgccagccct cccagtgctg tcattggccg ctacctgctg agcatcaggc 360 tttcctctcaccgcaaacac agcaaccgga ggctgggcga gtttgttctc cttttcaacc 420 catggtgtgcagaggacgat gtgtttctgg cctcagagga ggagagacag gagtacgtgc 480 tcagcgacagcggcatcatc ttccgaggcg tggagaagca catacgagcc cagggctgga 540 actacgggcagtttgaggag gacatcctga acatctgcct ctccatcctg gatcgaagcc 600 ccggtcaccaaaacaaccca gccaccgacg tgtcctgccg ccacaacccc atctacgtca 660 ccagggtcatcagtgccatg gtgaacagca acaacgaccg aggtgtggtg caaggacagt 720 ggcagggcaagtacggcggc ggcaccagcc cgctgcactg gcgcggcagc gtggccattc 780 tgcagaagtggctcaagggc aggtacaagc cagtcaagta cggccagtgc tgggtcttcg 840 ccggagtcctgtgcacagtc ctcaggtgct tggggatagc cacacgggtc gtgtccaact 900 tcaactcagcccacgacaca gaccagaacc tgagtgtgga caaatacgtg gactccttcg 960 ggcggaccctggaggacctg acagaagaca gcatgtggaa tttccatgtc tggaatgaga 1020 gctggtttgcccggcaggac ctaggcccct cttacaatgg ctggcaggtt ctggatgcca 1080 ccccccaggaggagagtgaa ggtgtgttcc ggtgcggccc agcctcagtc accgccatcc 1140 gcgagggtgatgtgcacctg gctcacgatg gccccttcgt gtttgcggag gtcaacgccg 1200 actacatcacctggctgtgg cacgaggatg agagccggga gcgtgtatac tcaaacacga 1260 agaagattgggagatgcatc agcaccaagg cggtgggcag tgactcccgc gtggacatca 1320 ctgacctctacaagtatccg gaagggtccc ggaaagagag gcaggtgtac agcaaggcgg 1380 tgaacaggctgttcggcgtg gaagcctctg gaaggagaat ctggatccgc agggctgggg 1440 gtcgctgtctctggcgtgac gacctcctgg agcctgccac caagcccagc atcgctggca 1500 agttcaaggtgctagagcct cccatgctgg gccacgacct gagactggcc ctgtgcttgg 1560 ccaacctcacctcccgggcc cagcgggtga gggtcaacct gagcggtgcc accatcctct 1620 atacccgcaagccagtggca gagatcctgc atgaatccca cgccgtgagg ctggggccgc 1680 aagaagagaagagaatccca attacaatat cttactctaa gtataaagaa gacctgacag 1740 aggacaagaagatcctgttg gctgccatgt gccttgtcac caaaggagag aagcttctgg 1800 tggagaaggacattactcta gaggacttca tcaccatcaa gcgtgcctac cctggagcct 1860 caggagagggcctcagtcca gtttgacatc accccctcca aaagtggccc aaggcagctg 1920 caggtggaccttgtaagccc tcacttcccg gacatcaagg gctttgtgat cgtccatgtg 1980 gccactgccaagtgatggat catgagggac tgagaggggt ggatttggcc cctgtcctcc 2040 tcctgcccattctttgtctc ttccacatgg gagccaggag gcctcagtta atcctgcctc 2100 aacct 2105144 626 PRT Homo sapiens 144 Met Thr Gln Gly Ile Arg Val Thr Lys Val AspTrp Gln Arg Ser Arg 1 5 10 15 Asn Gly Ala Ala His His Thr Gln Glu TyrPro Cys Pro Glu Leu Val 20 25 30 Val Arg Arg Gly Gln Ser Phe Ser Leu ThrLeu Glu Leu Ser Arg Ala 35 40 45 Leu Asp Cys Glu Glu Ile Leu Ile Phe ThrVal Glu Thr Gly Pro Arg 50 55 60 Ala Ser Glu Ala Leu His Thr Lys Ala ValPhe Gln Thr Ser Glu Leu 65 70 75 80 Glu Arg Gly Glu Gly Trp Thr Ala AlaArg Glu Ala Gln Met Glu Lys 85 90 95 Thr Leu Thr Val Ser Leu Ala Ser ProPro Ser Ala Val Ile Gly Arg 100 105 110 Tyr Leu Leu Ser Ile Arg Leu SerSer His Arg Lys His Ser Asn Arg 115 120 125 Arg Leu Gly Glu Phe Val LeuLeu Phe Asn Pro Trp Cys Ala Glu Asp 130 135 140 Asp Val Phe Leu Ala SerGlu Glu Glu Arg Gln Glu Tyr Val Leu Ser 145 150 155 160 Asp Ser Gly IleIle Phe Arg Gly Val Glu Lys His Ile Arg Ala Gln 165 170 175 Gly Trp AsnTyr Gly Gln Phe Glu Glu Asp Ile Leu Asn Ile Cys Leu 180 185 190 Ser IleLeu Asp Arg Ser Pro Gly His Gln Asn Asn Pro Ala Thr Asp 195 200 205 ValSer Cys Arg His Asn Pro Ile Tyr Val Thr Arg Val Ile Ser Ala 210 215 220Met Val Asn Ser Asn Asn Asp Arg Gly Val Val Gln Gly Gln Trp Gln 225 230235 240 Gly Lys Tyr Gly Gly Gly Thr Ser Pro Leu His Trp Arg Gly Ser Val245 250 255 Ala Ile Leu Gln Lys Trp Leu Lys Gly Arg Tyr Lys Pro Val LysTyr 260 265 270 Gly Gln Cys Trp Val Phe Ala Gly Val Leu Cys Thr Val LeuArg Cys 275 280 285 Leu Gly Ile Ala Thr Arg Val Val Ser Asn Phe Asn SerAla His Asp 290 295 300 Thr Asp Gln Asn Leu Ser Val Asp Lys Tyr Val AspSer Phe Gly Arg 305 310 315 320 Thr Leu Glu Asp Leu Thr Glu Asp Ser MetTrp Asn Phe His Val Trp 325 330 335 Asn Glu Ser Trp Phe Ala Arg Gln AspLeu Gly Pro Ser Tyr Asn Gly 340 345 350 Trp Gln Val Leu Asp Ala Thr ProGln Glu Glu Ser Glu Gly Val Phe 355 360 365 Arg Cys Gly Pro Ala Ser ValThr Ala Ile Arg Glu Gly Asp Val His 370 375 380 Leu Ala His Asp Gly ProPhe Val Phe Ala Glu Val Asn Ala Asp Tyr 385 390 395 400 Ile Thr Trp LeuTrp His Glu Asp Glu Ser Arg Glu Arg Val Tyr Ser 405 410 415 Asn Thr LysLys Ile Gly Arg Cys Ile Ser Thr Lys Ala Val Gly Ser 420 425 430 Asp SerArg Val Asp Ile Thr Asp Leu Tyr Lys Tyr Pro Glu Gly Ser 435 440 445 ArgLys Glu Arg Gln Val Tyr Ser Lys Ala Val Asn Arg Leu Phe Gly 450 455 460Val Glu Ala Ser Gly Arg Arg Ile Trp Ile Arg Arg Ala Gly Gly Arg 465 470475 480 Cys Leu Trp Arg Asp Asp Leu Leu Glu Pro Ala Thr Lys Pro Ser Ile485 490 495 Ala Gly Lys Phe Lys Val Leu Glu Pro Pro Met Leu Gly His AspLeu 500 505 510 Arg Leu Ala Leu Cys Leu Ala Asn Leu Thr Ser Arg Ala GlnArg Val 515 520 525 Arg Val Asn Leu Ser Gly Ala Thr Ile Leu Tyr Thr ArgLys Pro Val 530 535 540 Ala Glu Ile Leu His Glu Ser His Ala Val Arg LeuGly Pro Gln Glu 545 550 555 560 Glu Lys Arg Ile Pro Ile Thr Ile Ser TyrSer Lys Tyr Lys Glu Asp 565 570 575 Leu Thr Glu Asp Lys Lys Ile Leu LeuAla Ala Met Cys Leu Val Thr 580 585 590 Lys Gly Glu Lys Leu Leu Val GluLys Asp Ile Thr Leu Glu Asp Phe 595 600 605 Ile Thr Ile Lys Arg Ala TyrPro Gly Ala Ser Gly Glu Gly Leu Ser 610 615 620 Pro Val 625

1. A nucleotide sequence comprising at least a portion of the nucleotidesequence of FIG. 6A or FIG. 6B; a nucleotide sequence which hybridise tothe nucleotide sequence of FIG. 6A or FIG. 6B; a nucleotide sequencewhich is degenerate to the nucleotide sequence of FIG. 6A or FIG. 6B;all of which nucleotide sequences encode a polypeptide havingtransglutaminase activity.
 2. A nucleotide sequence according to claim 1consisting of the nucleotide sequence of FIG. 6A or FIG. 6B.
 3. Anucleotide sequence which hybridises under stringent conditions to thenucleotide sequence of FIG. 6A or FIG. 6B and which encodes apolypeptide having transglutaminase activity.
 4. A method of expressinga polypeptide comprising inserting a nucleotide sequence according toany preceding claim into a suitable host and expressing that nucleotidesequence in order to express a polypeptide having transglutaminaseactivity.
 5. A vector comprising a nucleotide sequence according to anyone of claims 1 to
 3. 6. A polypeptide having an amino acid sequencecomprising at least a portion of the amino acid sequence of FIG. 6A orFIG. 6B and which has transglutaminase activity.
 7. A polypeptideaccording to claim 6 which is at least 90% identical to the amino acidsequence of FIG. 6A or FIG. 6B and which encodes a polypeptide havingtransglutaminase activity.
 8. A polypeptide according to claim 6 or 7where the amino acid sequence differs from that given in FIG. 6A or FIG.6B by about 1 to 20 amino acid additions, deletions or substitutions. 9.A polypeptide according to any one of claims 6 to 8 comprising exon VIIthrough to exon X of the sequence shown in FIG. 6A or FIG. 6B.
 10. Acomposition comprising a polypeptide according to any one of claims 6 to9 suitable for use in cross-linking proteins.
 11. A compositioncomprising a polypeptide according to any one of claims 6 to 9 suitablefor use in a transamidation reaction on peptides and polypeptides.
 12. Adiagnostic method comprising detecting expression of a polypeptideaccording to any one of claims 6 to 9 in a subject or in cells derivedfrom a subject.
 13. An antibody directed against a polypeptide accordingto any one of claims 6 to
 9. 14. A method of gene therapy comprisingcorrecting mutations in a non wild type nucleotide sequencecorresponding to a nucleotide sequence of FIG. 6A or FIG. 6B.
 15. Anucleotide sequence comprising at least a portion of the nucleotidesequence of FIG. 10A or FIG. 10B; a nucleotide sequence which hybridiseto the nucleotide sequence of FIG. 10A or FIG. 10B; a nucleotidesequence which is degenerate to the nucleotide sequence of FIG. 10A orFIG. 10B; all of which nucleotide sequences encode a polypeptide havingtransglutaminase activity.
 16. A nucleotide sequence according to claim15 consisting of the nucleotide sequence of FIG. 10A or FIG. 10B.
 17. Anucleotide sequence which hybridises under stringent conditions to thenucleotide sequence of FIG. 10A or FIG. 10B and which encodes apolypeptide having transglutaminase activity.
 18. A method of expressinga polypeptide comprising inserting a nucleotide sequence according toany one of claims 15 to 17 into a suitable host and expressing thatnucleotide sequence in order to express a polypeptide havingtransglutaminase activity.
 19. A vector comprising a nucleotide sequenceaccording to any one of claims 15 to
 17. 20. A polypeptide having anamino acid sequence comprising at least a portion of the amino acidsequences of FIG. 10A or FIG. 10B and which has transglutaminaseactivity.
 21. A polypeptide according to claim 20 which is at least 90%identical to the amino acid sequences of FIG. 10A or FIG. 10B and whichencodes a polypeptide having transglutaminase activity.
 22. Apolypeptide according to claim 20 or 21 wherein the amino acid sequencediffers from that given in FIG. 10A or FIG. 10B by about 1 to 20 aminoacid additions, deletions or substitutions.
 23. A polypeptide accordingto any one of claims 20 to 22 comprising exons II through to exon IV ofthe sequence shown in FIG. 10A or FIG. 10B.
 24. A polypeptide accordingto any one of claims 20 to 22 comprising exon X through to exon XII ofthe sequence shown in FIG. 10A or FIG. 10B.
 25. A composition comprisinga polypeptide according to any one of claims 20 to 24 suitable for usein cross-linking proteins.
 26. A composition comprising a polypeptideaccording to any one of claims 20 to 24 suitable for use in atransamidation reaction on peptides and polypeptides.
 27. A diagnosticmethod comprising detecting expression of a polypeptide according to anyone of claims 20 to 24 in a subject or in cells derived from a subject.28. An antibody directed against a polypeptide according to any one ofclaims 20 to
 24. 29. A method of gene therapy comprising correctingmutations in a non-wild type nucleotide sequence corresponding to thenucleotide sequence of FIG. 10A or FIG. 10B.
 30. A method of diagnosisof autoimmune disease comprising taking a sample from a subject andtesting that sample for the presence of a transglutaminase encoded bythe nucleotide sequences of FIG. 6A, FIG. 6B, FIG. 10A or FIG. 10B, orportions thereof.
 31. A method according to claim 30 wherein theautoimmune disease to be diagnosed is selected from Addison's disease,AI haemolytic anaemia, AI thrombocytopenic purpura, AI thyroid diseases,atrophic gastritis—pernicious anaemia, Chron's disease, colitisulcerosa, Goodpasture syndrome, IgA nephropathy or IgAglomerulonephritis, myasthenia gravis, partial lipodystrophy,polymyositis, primary biliary cirrhosis, primary sclerosing cholangitis,progressive systemic sclerosis, recurrent pericarditis, relapsingpolychondritis, rheumatoid arthritis, rheumatism, sarcoidosis, Sjögren'ssyndrome, SLE, splenic atrophy, type I (insulin-dependent) diabetesmellitus, diabetis mellitus, Wegener granulomatosis, ulcerative colitis,vasculitis (both systemic and cutaneous), vitiligo.
 32. A competitiveprotein binding assay for the differential diagnosis of autoimmunediseases comprising the detection of antibodies against thetransglutaminase encoded by the nucleotide sequences of FIG. 6A, FIG.6B, FIG. 10A or FIG. 10B, or portions thereof.
 33. A competitive proteinbinding assay according to claim 32 comprising non-endogenoustransglutaminase TG_(Z) or TG_(Y), or both, as a competitive antigen.34. Competitive protein binding assay according to claim 33, wherein thebinding assay is a competitive immunoassay selected from RIA, EIA/ELISA,LiA and FiA.